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Product listing: CYLD Antibody, UniProt ID Q9NQC7 #4495 to Autophagy Antibody Sampler Kit, UniProt ID O94817 #4445

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: CYLD is a cytoplasmic deubiquitinating enzyme encoded by a tumor suppressor gene altered in individuals diagnosed with cylindromatosis, a genetic condition characterized by benign tumors of skin appendages (1,2). Functional CYLD deubiquitinase regulates inflammation and cell proliferation by down regulating NF-κB signaling through removal of ubiquitin chains from several NF-κB pathway proteins (3,4). CYLD is a negative regulator of proximal events in Wnt/β-catenin signaling and is a critical regulator of natural killer T cell development (5,6). The transcription factor Snail can inhibit CYLD expression, resulting in melanoma cell proliferation (7).

$303
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Dynamin-related protein 1 (DRP1) is a member of the dynamin superfamily of GTPases. Members of this family have diverse cellular functions including vesicle scission, organelle fission, viral resistance, and intracellular trafficking (reviewed in 1). DRP1 affects mitochondrial morphology and is important in mitochondrial and peroxisomal fission in mammalian cells (2-5). The yeast ortholog of DRP1 clusters into a spiral-shaped structure on the mitochondrial membrane at the site of fission (reviewed in 6), and this structure is likely conserved in mammalian cells (3). The division of the mitochondria, which is required for apoptosis, as well as normal cell growth and development is controlled, in part, by the phosphorylation of DRP1 at Ser616 by Cdk1/cyclin B and at Ser637 by protein kinase A (PKA) (reviewed in 6). When phosphorylated at Ser616, DRP1 stimulates mitochondrial fission during mitosis. Conversely, fission is inhibited when DRP1 is phosphorylated at Ser637 (reviewed in 6). Dephosphorylation at Ser637 by calcineurin reverses this inhibition (7). In addition to phosphorylation, sumoylation of DRP1 is also an enhancer of mitochondrial fission (8). Balancing fission and fusion events is essential for proper mitochondrial function. Research studies have demonstrated mitochondrial defects in a variety of neurodegenerative diseases including Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease (reviewed in 6).

$108
250 PCR reactions
500 µl
SimpleChIP® Human MYT-1 Exon 1 Primers contain a mix of forward and reverse PCR primers that are specific to exon 1 of the human myelin transcription factor 1 (MYT-1) gene. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in SYBR® Green quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003 and ChIP-validated antibodies from Cell Signaling Technology®. The MYT-1 gene is repressed in most cell types and is bound by polycomb group proteins such as Ezh2, Suz12, Rnf2 and Bmi-1. MYT-1 exon 1 is also enriched for histone H3 Lys27 tri-methylation and histone H2A Lys119 ubiquitylation.
REACTIVITY
Human

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to either identify multiple proteins associated with a specific region of the genome or to identify the many regions of the genome bound by a particular protein (3-6). ChIP can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors, and DNA repair proteins. When performing the ChIP assay, cells are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. Fragmented chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or quantitative real-time PCR are often used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing (ChIP-Seq), or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8). SimpleChIP® primers have been optimized for amplification of ChIP-isolated DNA using real-time quantitative PCR and provide important positive and negative controls that can be used to confirm a successful ChIP experiment.

$303
100 µl
APPLICATIONS
REACTIVITY
Rat

Application Methods: Western Blotting

Background: Tpl2 (tumor progression locus 2), also known as COT (cancer osaka thyroid), is a serine/threonine kinase expressed primarily in hematopoietic tissues, lung and liver (1). Over-expression of Tpl2 potentiates MAP kinase pathways through MEK1 and SEK1, as well as through MKK6 and MEK5 (2,3). Tpl2 is also engaged in NF-κB activation through NF-κB inducing kinase (NIK), or by inducing phosphorylation and degradation of the NF-κB precusor, p105 NF-κB1 (4,5). Ser400 of Tpl2 is phosphorylated in an Akt-dependent manner. This phosphorylation is required for Tpl2-induced NF-κB-dependent transcription (6). Tpl2 also activates caspase-3 by promoting the assembly of a protein complex of Apaf1 (apoptotic protease-activating factor 1), caspase-9, Tpl2, adaptor protein Tvl1 and procaspase-3 (7).

$108
250 PCR reactions
500 µl
SimpleChIP® Human MyoD1 Exon 1 Primers contain a mix of forward and reverse PCR primers that are specific to the exon 1 region of the human myoblast determination protein 1 gene. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in SYBR® Green quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003 and ChIP-validated antibodies from Cell Signaling Technology®. The MyoD1 gene is expressed primarily in muscle cells and is inactive in most other cell types, where it is enriched for histone modifications associated with repressed genes, such as histone H3 Lys9 di-methylation and Lys27 tri-methylation.
REACTIVITY
Human

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to either identify multiple proteins associated with a specific region of the genome or to identify the many regions of the genome bound by a particular protein (3-6). ChIP can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors, and DNA repair proteins. When performing the ChIP assay, cells are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. Fragmented chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or quantitative real-time PCR are often used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing (ChIP-Seq), or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8). SimpleChIP® primers have been optimized for amplification of ChIP-isolated DNA using real-time quantitative PCR and provide important positive and negative controls that can be used to confirm a successful ChIP experiment.

The RNAi Machinery Antibody Sampler Kit provides an economical means to analyze proteins associated with endogenous RNA interference. The kit contains enough primary and secondary antibodies to perform two western blot experiments.
$108
250 PCR reactions
500 µl
SimpleChIP® Human α Satellite Repeat Primers contain a mix of forward and reverse PCR primers that are specific to the human α satellite repeat element. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in SYBR® Green quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003 and ChIP-validated antibodies from Cell Signaling Technology®. The human α satellite repeat element is found in pericentromeric regions of chromosomes and shows hallmarks of heterochromatin including histone H3 Lys9 di- and tri-methylation, as well as HP1 binding.
REACTIVITY
Human

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to either identify multiple proteins associated with a specific region of the genome or to identify the many regions of the genome bound by a particular protein (3-6). ChIP can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors, and DNA repair proteins. When performing the ChIP assay, cells are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. Fragmented chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or quantitative real-time PCR are often used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing (ChIP-Seq), or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8). SimpleChIP® primers have been optimized for amplification of ChIP-isolated DNA using real-time quantitative PCR and provide important positive and negative controls that can be used to confirm a successful ChIP experiment.

$327
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649.
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat, Zebrafish

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry)

Background: Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: THEMIS is a recently identified protein found to be critical for T cell development (1-5). It contains two amino-terminal globular domains, a nuclear localization signal, and a carboxy-terminal proline-rich sequence with homology to SH3 binding domains (2,3). THEMIS is detected in both the cytoplasm and the nucleus (2). It is expressed at low levels in mature T cells and during thymocyte development, with expression peaking in CD4+CD8+ double positive cells (1-5). THEMIS is tyrosine phosphorylated downstream of TCR signaling and associates with the adaptor GRB2 and possibly other SH3 domain-containing proximal TCR signaling molecules (1-5). Mice lacking THEMIS have a defect in positive selection that results in decreased numbers of both single positive thymocytes and mature T cells (1-5).

$303
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: PTPα (PTPRA) is a transmembrane receptor tyrosine phosphatase implicated in the regulation of Src family kinases during the G2 to mitosis entry point. Two identified splice variants differ in the size of the extracellular region; the shorter form appears to be ubiquitously expressed while the larger protein is more limited in distribution (1). The cytoplasmic region of PTPα contains two putative catalytic domains. One phosphatase domain (D1) exhibits catalytic activity while the other (D2) may regulate phosphatase activity by allowing receptor dimer formation (2,3). PTPα is a physiological regulator of Src and Src family kinases (4). Constitutive phosphorylation of the carboxy-terminal Tyr789 of PTPα is essential for dephosphorylation of Src at Tyr527. Phosphorylation of PTPα at this residue also allows binding of the Grb2 inhibitor, restricting PTPα activation of Src (5,6). PKC-mediated phosphorylation of the PTP at Ser180 and Ser204 also increases PTPα phosphatase activity (7).

$108
250 PCR reactions
500 µl
SimpleChIP® Human GAPDH Intron 2 Primers contain a mix of forward and reverse PCR primers that are specific to intron 2 of the human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in SYBR® Green quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003 and ChIP-validated antibodies from Cell Signaling Technology®. The GAPDH gene is actively transcribed in all cell types and it is highly enriched for histone modifications associated with active transcription, such as histone H3 Lys4 tri-methylation and general histone acetylation. The intron 2 region is highly enriched for histone modifications found in the body of active genes, such as histone H3 Lys36 tri-methylation and histone H2B Lys120 ubiquitination.
REACTIVITY
Human

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to either identify multiple proteins associated with a specific region of the genome or to identify the many regions of the genome bound by a particular protein (3-6). ChIP can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors, and DNA repair proteins. When performing the ChIP assay, cells are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. Fragmented chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or quantitative real-time PCR are often used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing (ChIP-Seq), or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8). SimpleChIP® primers have been optimized for amplification of ChIP-isolated DNA using real-time quantitative PCR and provide important positive and negative controls that can be used to confirm a successful ChIP experiment.

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: ABCG2 (BCRP1/ABCP/MXR) is a member of the ATP-binding cassette transporter family that functions as ATP-dependent transporters for a wide variety of chemical compounds and are associated with drug-resistance in cancer cells (1-6). ABCG2 is a heavily glycosylated transmembrane protein with six transmembrane spanning regions consistent with it functioning as a half-transporter. The ABC family can exist as either full-length transporters or as half-transporters that form functional transporters through homo- or heterodimerization. High expression of ABCG2 was found in placenta as well as cell lines selected for resistance to a number of chemotherapeutic drugs, including mitoxantrone, doxorubicin, topotecan and flavopiridol. In rodents, the highest expression of ABCG2 was found in kidney (8). ABCG2 expression has also been observed in stem cell populations, particularly in hematopoietic and neuronal stem cells and is downregulated with differentiation (9-12).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: Scribble (Scrib) was originally identified in a genetic screen in Drosophila along with cell polarity determinants Discs Large (Dlg) and Lethal giant larvae (Lgl). Drosophila mutants homozygous for these genes share similar phenotypes, including the loss of apicobasal cell polarity and neoplastic tissue overgrowth. These phenotypic similarities suggest that these three proteins function in a common pathway important for establishing and maintaining apicobasal polarity in epithelial cells (1,2). Scribble contains many leucine-rich repeats and PDZ domains important for localizing scribble to adherens junctions and basolateral regions of mammalian epithelial cells (3). Scribble reportedly binds β-catenin, APC, E-cadherin and the E6 protein from high-risk virus type of HPV through a short motif important for E6-induced cell transformation (4-8). Overexpression of scribble inhibits transformation of rodent epithelial cells by HPV E6/7 proteins (8).

$134
20 µl
$336
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Chromatin IP, Flow Cytometry, Immunofluorescence (Immunocytochemistry), Immunoprecipitation, Western Blotting

Background: The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: Hematopoietic progenitor kinase 1(HPK1) is a member of the germinal center kinases that is predominantly expressed in hematopoietic cells. HPK1 is a key component linking T or B cell receptors to SAPK/JNK and IkappaB kinase pathways in lymphocytes (1-3). Through its proline-rich motif, HPK1 associates with multiple SH3 domain-containing adaptor proteins, including GRB2, Nck, Crk, SLP-76, and BLNK. Phosphorylation of Tyr379 by Syk is necessary for HPK1 association with BLNK and SLP-76, as well as for HPK1 activity (4,5). Caspase cleavage of HPK1 also modulates the the biological function of HPK1 (3,6,7).

$108
250 PCR reactions
500 µl
SimpleChIP® Human GAPDH Promoter Primers contain a mix of forward and reverse PCR primers that are specific to a region of the human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in SYBR® Green quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9002 and #9003 and ChIP-validated antibodies from Cell Signaling Technology®. The GAPDH gene is actively transcribed in all cell types and its promoter is highly enriched for histone modifications associated with active transcription, such as histone H3 Lys4 tri-methylation and general histone acetylation. This gene promoter shows very low levels of histone modifications associated with heterochromatin, such as histone H3 Lys9 or Lys27 tri-methylation.
REACTIVITY
Human

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to either identify multiple proteins associated with a specific region of the genome or to identify the many regions of the genome bound by a particular protein (3-6). ChIP can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors, and DNA repair proteins. When performing the ChIP assay, cells are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. Fragmented chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or quantitative real-time PCR are often used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing (ChIP-Seq), or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8). SimpleChIP® primers have been optimized for amplification of ChIP-isolated DNA using real-time quantitative PCR and provide important positive and negative controls that can be used to confirm a successful ChIP experiment.

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: The mTORC1 kinase complex is a critical regulator of cell growth (1,2). Its activity is modulated by energy levels, growth factors, and amino acids via signaling through Akt, MAPK, and AMPK pathways (3,4). Recent studies found that the four related GTPases, RagA, RagB, RagC, and RagD, interact with raptor within the mTORC1 complex (1,2). These interactions are both necessary and sufficient for mTORC1 activation in response to amino acid signals (1,2).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Bif-1/SH3GLB1/Endophilin-B1 is a member of the endophilin B family that was originally identified as a Bax binding protein through yeast two-hybrid screening (1,2). Bif-1 does not have significant homology to other Bcl-2 family members, but rather contains an N-terminal Bin-Amphiphysin-Rvs (BAR) domain, typically involved in membrane dynamics, and a C-terminal SH3 domain. Overexpression of Bif-1 promotes Bax conformational change and apoptosis (2,3). Likewise, loss of Bif-1 inhibits Bax and Bak activation, cytochrome c release, and caspase activation (3). Bif-1 is localized to membranes of intracellular organelles and has been suggested to play a role in membrane dynamics, including that during autophagy. Bif-1 directly binds to UVRAG, forming a complex with Beclin-1, resulting in increased PI3-kinase class III/Vps34 activity required for autophagosome maturation (4). Inhibition of GSK-3β, as seen during nutrient deprivation, results in increased expression of Bif-1, and can contribute to autophagic cell death (5). Research studies have shown that loss of Bif-1 promotes tumorigenesis, and decreased expression of Bif-1 has been noted in several cancer types (6-11).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunofluorescence (Frozen), Immunohistochemistry (Paraffin), Western Blotting

Background: The cytoskeleton consists of three types of cytosolic fibers: microtubules, microfilaments (actin filaments), and intermediate filaments. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. γ-tubulin is required to nucleate polymerization of tubulin subunits to form microtubule polymers. Many cell movements are mediated by microtubule action, including the beating of cilia and flagella, cytoplasmic transport of membrane vesicles, chromosome alignment during meiosis/mitosis, and nerve-cell axon migration. These movements result from competitive microtubule polymerization and depolymerization or through the actions of microtubule motor proteins (1).β3-tubulin (TUBB3) is one of six β-tubulin isoforms and is expressed highly during fetal and postnatal development (axon guidance and maturation) (2). Its expression levels decrease in the adult central nervous system (CNS) but remain high in the peripheral nervous system (PNS) (3). Microtubules enriched in β3-tubulin are more dynamic than those composed of other β-tubulin isoforms (4). Research studies have shown that mutations in the β3-tubulin gene TUBB3 cause ocular motility defects and other nervous system disorders. Furthermore, β3-tubulin is present in neoplastic but not in normal differentiated glial cells. Thus, β3-tubulin is a great neuronal marker (5).

$122
20 µl
$293
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Keratins (cytokeratins) are intermediate filament proteins that are mainly expressed in epithelial cells. Keratin heterodimers composed of an acidic keratin (or type I keratin, keratins 9 to 23) and a basic keratin (or type II keratin, keratins 1 to 8) assemble to form filaments (1,2). Keratin isoforms demonstrate tissue- and differentiation-specific profiles that make them useful as research biomarkers (1). Research studies have shown that mutations in keratin genes are associated with skin disorders, liver and pancreatic diseases, and inflammatory intestinal diseases (3-6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: HOP, also known as stress-induced phospho protein 1 (STIP), is a co-chaperone to the major heat shock proteins, Hsp70 and Hsp90, and appears in early receptor complexes (1,2). Through mutual binding to both Hsp70 and Hsp90, Hop functions as an adaptor that can integrate Hsp70 and Hsp90 interactions (3,4). HOP is an abundant and highly conserved protein which is composed of three tetratricopeptide repeat (TPR) domains (TPR1, TPR2a and TPR2b) and two DP repeat domains (DP1 and DP2), whose function has not been fully resolved (5).

$260
100 µl
APPLICATIONS
REACTIVITY
Bovine, Hamster, Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Ran is a small GTPase of the Ras family that plays a central role in the spacial and temperal organization of eukaryotic cells. During interphase, Ran-GDP localizes to the cytoplasm and Ran-GTP to the nucleus. This polarized localization of Ran ensures its role in nuclear transport (1). During mitosis, Ran-GTP is chromatin associated, where it promotes spindle assembly and nuclear envolope formation (1,2). In S phase, Ran-GTP associates with and inhibits MCM helicase, ensuring precise chromosomal DNA duplication during the cell cycle (3).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: AMP-activated protein kinases (AMPKs) constitute a serine/threonine protein kinase family, which is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (1). AMPKα1, AMPKα2, MELK and SNARK are the catalytic subunits in the family (2). Recently, AMPK-related protein kinase ARK5 was identified, which shares 84% similarity with the sequence of the catalytic domain of SNARK (2). In vitro, Akt phosphorylates ARK5 at Ser600. This phosphorylation activates ARK5, which in turn results in the increased phosphorylation of the SAMS peptide, an AMPK consensus substrate (2). In vivo experiments showed that Akt-activated ARK5 is critical for the survival of cells under glucose starvation (2). Furthermore, studies also linked ARK5 to tumor invasion (3, 4). Overexpressed ARK5 leads to higher tumor growth rate (3). For example, the overexpression of ARK5 in pancreatic tumor cell line PANC-1 significantly elevates its metastasis in the liver (4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Nervous system nuclear protein induced by axotomy 1 (Nna1, AGTPBP1 or cytosolic carboxypeptidase-1) is related to zinc carboxypeptidases and contains an ATP/GTP binding motif, a basic and a bipartite nuclear localization signal. Nna1 expression is rapidly induced in motor neurons following axotomy and down-regulated following reinnervation, which is consistent with abundant Nna1 expression in differentiating neurons and the absence of Nna1 in CNS proliferative zones (1). Furthermore, Purkinje cell degeneration is characterized by adult onset neurodegeneration resulting from Nna1 gene mutations, implicating Nna1 in mechanisms common to degeneration and regeneration (2).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: CTP:phosphocholine cytidylyltransferase (CCT) is a critical enzyme that regulates the CDP-choline pathway for the biosynthesis of phosphatidylcholine. Three distinct CCT isoforms are found in mammals, including CCTα, CCTβ2, and CCTβ3 (1,2). CCTα is the major isoform that is expressed in most tissues (3). CCTα is essential in the synthesis and secretion of surfactant by alveolar epithelial cells and is important in maintaining the phosphatidylcholine level that regulates lipoprotein assembly and secretion in hepatocytes (4,5). CCTα is a major component in membrane biogenesis during cytokine secretion by stimulated macrophages (6). Monoubiquitination of CCTα prevents it from entering the nucleus and leads to its degradation by lysosome (7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Caspase-4 (TX/ICH-2/ICErelII) is a member of the caspase family of proteases that play a key role in the execution of apoptosis and activation of inflammatory cytokines (1-3). Expression of caspase-4 has been observed in most tissues except brain, with highest levels in placenta, lung, spleen, and peripheral blood lymphocytes (PBL). Caspase-4 was originally found to contribute to Fas-mediated apoptosis (4). Several caspases (including caspase-4, caspase-5, and mouse caspase-11 and -12) are most closely related to caspase-1 and are capable of inducing apoptosis when over-expressed but are better characterized in the proteolytic activation of inflammatory cytokines (5). Caspase-4 associates with TRAF6 and is involved in the LPS inducible production of inflammatory cytokines IL-8 and MIP1 in THP-1 cells (6). While caspase-4 and mouse caspase-12 localize to the endoplasmic reticulum (ER) and may be activated by drugs that induce ER-stress (7), at least one study suggests that caspase-4 and caspase-12 are not essential for the ER-stress induced apoptosis (8).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: The cohesin complex consists of a heterodimer between SMC1 (SMC1A or B) and SMC3, bound by additional RAD21 and STAG proteins (STAG1, 2, or 3) (1,2). These proteins form a ring-like structure that mediates the cohesion of two sister chromatids after DNA replication in S phase (1,2). RAD21 and STAG2 are phosphorylated by Polo-like kinase (PLK) during prophase, which leads to the dissociation of cohesin complexes from the chromosome arms; however, cohesin remains bound to centromeres until anaphase (3,4). RAD21 is cleaved by separin/ESPL1 in anaphase, which leads to dissociation of the remaining cohesin from centromeres, enabling sister chromatids to segregate during mitosis (5). RAD21 is also cleaved by caspase-3 and caspase-7 during apoptosis, resulting in a 64 kDa carboxy-terminal cleavage product that translocates to the cytoplasm and may help to trigger apoptosis (6,7). In addition to mediating cohesion of sister chromatids, the cohesin complex plays important roles in gene regulation and DNA repair, as SMC1 and SMC3 are both phosphorylated by ATM and ATR kinases upon DNA damage (1,2).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: Alpha-fetoprotein (AFP) is a 65 kDa glycoprotein found in the mammalian fetal liver, yolk sac, and GI tract. While AFP expression in adult cells is low, it is aberrantly expressed in adult liver cancer cells (1,2). The tumor suppressor gene p53 and β-catenin are both involved in the regulation of AFP expression. In normal adult cells, p53 binds to the repressor region of the AFP gene, thereby blocking transcription. Mutations in both p53 and β-catenin are associated with aberrant expression of AFP. Research studies have shown that elevated serum AFP levels are predictive of hepatocellular carcinoma (3).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunofluorescence (Immunocytochemistry), Immunoprecipitation, Western Blotting

Background: Multi-drug resistance protein 2 (MRP2), also known as cMRP, cMOAT, and ABCC2, is an ATP binding cassette (ABC) transporter and part of the multi-drug resistance (MRP) family (1,2). The MRP proteins are membrane proteins that function as organic anion pumps involved in the cellular removal of cancer drugs (2). MRP2 is associated with resistance to a number of cancer drugs, such as cisplatin, etoposide, doxorubicin, and methotrexate (3-5). MRP2 is predominately expressed on the apical membranes in the liver (6-9) and kidney proximal tubules (10). It is responsible for the ATP-dependent secretion of bilirubin glucuronides and other organic anions from hepatocytes into the bile, a process important for the excretion of endogenous and xenobiotic substances. Loss of MRP2 activity is the cause of Dubin-Johnson syndrome, an autosomal recessive disorder characterized by defects in the secretion of anionic conjugates and the presence of melanin like pigments in hepatocytes (11-13).

The Autophagy Antibody Sampler Kit provides an economical means to investigate the molecular machinery of autophagy within the cell. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.

Background: Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegeneration, infection, and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes. Formation of the autophagosome involves a ubiquitin-like conjugation system in which Atg12 is covalently bound to Atg5 and targeted to autophagosome vesicles (4-6). This conjugation reaction is mediated by the ubiquitin E1-like enzyme Atg7 and the E2-like enzyme Atg10 (7,8).