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Product listing: AS160 Antibody, UniProt ID O60343 #2447 to ASF1B Antibody, UniProt ID Q9NVP2 #2769

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: Insulin is a major hormone controlling critical energy functions, such as glucose and lipid metabolism. Insulin binds to and activates the insulin receptor (IR) tyrosine kinase, which phosphorylates and recruits adaptor proteins. The signaling pathway initiated by insulin and its receptor stimulates glucose uptake in muscle cells and adipocytes through translocation of the Glut4 glucose transporter from the cytoplasm to the plasma membrane (1). A 160 kDa substrate of the Akt Ser/Thr kinase (AS160, TBC1D4) is a Rab GTPase-activating protein that regulates insulin-stimulated Glut4 trafficking. AS160 is expressed in many tissues including brain, kidney, liver, and brown and white fat (2). Multiple Akt phosphorylation sites have been identified on AS160 in vivo, with five sites (Ser318, Ser570, Ser588, Thr642, and Thr751) showing increased phosphorylation following insulin treatment (2,3). Studies using recombinant AS160 demonstrate that insulin-stimulated phosphorylation of AS160 is a crucial step in Glut4 translocation (3) and is reduced in some patients with type 2 diabetes (4). The interaction of 14-3-3 regulatory proteins with AS160 phosphorylated at Thr642 is a necessary step for Glut4 translocation (5). Phosphorylation of AS160 by AMPK is involved in the regulation of contraction-stimulated Glut4 translocation (6).

$303
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Transcription factor EB (TFEB) is a member of the Myc-related, bHLH leucine-zipper family of transcription factors that drives the expression of a network of genes known as the Coordinated Lysosomal Expression and Regulation (CLEAR) network (1,2). TFEB specifically recognizes and binds regulatory sequences within the CLEAR box (GTCACGTGAC) of lysosomal and autophagy genes, resulting in the up-regulated expression of genes involved in lysosome biogenesis and function, and regulation of autophagy (1,2). TFEB is activated in response to nutrient deprivation, stimulating translocation to the nucleus where it forms homo- or heterooligomers with other members of the microphthalmia transcription factor (MiTF) subfamily and resulting in up-regulation of autophagosomes and lysosomes (3-5). Recently, it has been shown that TFEB is a component of mammalian target of rapamycin (mTOR) complex 1 (mTORC1), which regulates the phosphorylation and nuclear translocation of TFEB in response to cellular starvation and stress (6-9). During normal growth conditions, TFEB is phosphorylated at Ser211 in an mTORC1-dependent manner. Phosphorylation promotes association of TFEB with 14-3-3 family proteins and retention in the cytosol. Inhibition of mTORC1 results in a loss of TFEB phosphorylation, dissociation of the TFEB/14-3-3 complex, and rapid transport of TFEB to the nucleus where it increases transcription of CLEAR and autophagy genes (10). TFEB has also been shown to be activated in a nutrient-dependent manner by p42 MAP kinase (Erk2). TFEB is phosphorylated at Ser142 by Erk2 in response to nutrient deprivation, resulting in nuclear localization and activation, and indicating that pathways other than mTOR contribute to nutrient sensing via TFEB (3).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: Niemann-Pick C1-like 1 (NPC1L1) is a transmembrane protein that plays a critical role in cholesterol absorption (1). It is highly expressed in small intestine and localized along the brush border in both human and mouse epithelial cells (2,3). NPC1L1 mediates cholesterol uptake via vesicular endocytosis (4). Ezetimibe, a potent cholesterol absorption inhibitor used to treat hypercholesterolemia (5), inhibits cholesterol uptake by preventing NPC1L1 from incorporating into clathrin-coated vesicles (4).

$305
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 555 fluorescent dye and tested in-house for immunofluroescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated α-Tubulin (11H10) Rabbit mAb #2125.
APPLICATIONS
REACTIVITY
Bovine, D. melanogaster, Human, Monkey, Mouse, Pig, Rat, Zebrafish

Application Methods: Immunofluorescence (Immunocytochemistry)

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).

$260
100 µl
APPLICATIONS
REACTIVITY
Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2, and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as de novo methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither de novo nor maintenance DNA methylation.DNMT3L is a catalytically inactive regulatory factor for the DNMT3A and DNMT3B de novo methyltransferases that is expressed at low levels in embryonic stem cells, testis, ovaries, and thymus (1,2). These de novo methyltransferases consist of a heterotetrameric complex containing two molecules of DNMT3L, and either two molecules of DNMT3A or DNMT3B (3). DNMT3L contains an amino-terminal ATRX-DNMT3-DNMT3L (ADD) domain and a carboxy-terminal methyltransferase-like domain (4-7). The methyltransferase-like domain binds to DNMT3A and DNMT3B to stimulate catalytic activity by increasing the binding of S-adenosylmethionine and DNA (4,5). The ADD domain recruits the methyltransferase complex to transcriptionally inactive regions of the genome by binding to unmethylated histone H3 Lys4 (6,7).

$108
250 PCR reactions
500 µl
SimpleChIP® Mouse RPL30 Intron 2 Primers contain a mix of forward and reverse PCR primers that are specific to intron 2 of the mouse RPL30 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 RPL30 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 and Lys27 tri-methylation.
REACTIVITY
Mouse

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: AMPA- (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid), kainate-, and NMDA- (N-methyl-D-aspartate) receptors are the three main families of ionotropic glutamate-gated ion channels. AMPA receptors (AMPARs) are comprised of four subunits (GluR 1-4), which assemble as homo- or hetero-tetramers to mediate the majority of fast excitatory transmissions in the central nervous system. AMPARs are implicated in synapse formation, stabilization, and plasticity (1). In contrast to GluR 2-containing AMPARs, AMPARs that lack GluR 2 are permeable to calcium (2). Post-transcriptional modifications (alternative splicing, nuclear RNA editing) and post-translational modifications (glycosylation, phosphorylation) result in a very large number of permutations, fine-tuning the kinetic properties of AMPARs. Research studies have implicated activity changes in AMPARs in a variety of diseases including Alzheimer’s, amyotrophic lateral sclerosis (ALS), stroke, and epilepsy (1).

$303
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: The Forkhead family of transcription factors is involved in tumorigenesis of rhabdomyosarcoma and acute leukemias (1-3). Within the family, three members (FoxO1, FoxO4, and FoxO3a) have sequence similarity to the nematode orthologue DAF-16, which mediates signaling via a pathway involving IGFR1, PI3K, and Akt (4-6). Active forkhead members act as tumor suppressors by promoting cell cycle arrest and apoptosis. Increased expression of any FoxO member results in the activation of the cell cycle inhibitor p27 Kip1. Forkhead transcription factors also play a part in TGF-β-mediated upregulation of p21 Cip1, a process negatively regulated through PI3K (7). Increased proliferation results when forkhead transcription factors are inactivated through phosphorylation by Akt at Thr24, Ser256, and Ser319, which results in nuclear export and inhibition of transcription factor activity (8). Forkhead transcription factors can also be inhibited by the deacetylase sirtuin (SirT1) (9).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: ASF1 was first identified in S. cerevisiae based on its ability to de-repress transcriptional silencing when overexpressed (1). While only one gene exists in yeast and Drosophila, mammalian cells contain the two highly homologous ASF1A and ASF1B genes (2). ASF1A and ASF1B function as histone chaperones, delivering histone H3/H4 dimers to CAF-1 or HIRA histone deposition complexes to facilitate replication-coupled and replication-independent nucleosome assembly on DNA (2-5). Both ASF1A and ASF1B bind to CAF-1, but only ASF1A binds to HIRA (5). In addition to playing a role in DNA replication and gene silencing, ASF1 functions in DNA damage repair, genome stability and cellular senescence. Deletion of ASF1 in yeast and Drosophila confers sensitivity to various DNA damaging agents and inhibitors of DNA replication, increases genomic instability and sister chromatid exchange, and activates the DNA damage checkpoint (6-8). Depletion of both ASF1A and ASF1B in mammalian cells results in the accumulation of cells in S phase, increased phosphorylation of H2A.X, centrosome amplification and apoptosis (9,10). ASF1A is required for the formation of senescence-associated heterochromatin foci (SAHF), with overexpression of ASF1A inducing senescence in primary cells (4). Both ASF1A and ASF1B are phosphorylated in S phase by the Tousled-like kinases TLK1 and TLK2, and are dephosphorylated when TLK1 and TLK2 are inactivated by Chk1 kinase in response to replicative stress (11,12). The function of ASF1 phosphorylation is not yet understood.

$499
96 assays
1 Kit
The FastScan™ Phospho-Rb (Ser807/811) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Rb when phosphorylated at Ser807/811. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with phospho-Rb (Ser807/811) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of phospho-Rb (Ser807/811). Antibodies in kit are custom formulations specific to kit.
REACTIVITY
Human, Monkey

Background: The retinoblastoma tumor suppressor protein Rb regulates cell proliferation by controlling progression through the restriction point within the G1-phase of the cell cycle (1). Rb has three functionally distinct binding domains and interacts with critical regulatory proteins including the E2F family of transcription factors, c-Abl tyrosine kinase, and proteins with a conserved LXCXE motif (2-4). Cell cycle-dependent phosphorylation by a CDK inhibits Rb target binding and allows cell cycle progression (5). Rb inactivation and subsequent cell cycle progression likely requires an initial phosphorylation by cyclin D-CDK4/6 followed by cyclin E-CDK2 phosphorylation (6). Specificity of different CDK/cyclin complexes has been observed in vitro (6-8) and cyclin D1 is required for Ser780 phosphorylation in vivo (9).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunohistochemistry (Paraffin), Western Blotting

Background: Insulin-like growth factor II (IGF-II) receptor, also widely known as cation-independent mannose 6-phosphate receptor (CI-M6PR), is a multifunctional type I transmembrane glycoprotein that participates in the internalization of mannose-6-phosphate modified hydrolases and IGF-II from the plasma membrane (1,2). In the absence of ligands, IGF-II receptor is constitutively endocytosed from the cell surface to accumulate in the Golgi apparatus (3). In the presence of ligands, the receptor transports the mannose-6-phosphate modified hydrolases to acidified endosomes and lysosomes (4). The ligand-free receptor is then transported back to the Golgi compartment or the cell surface (4). In several research studies, IGF-II receptor has been recognized as a tumor suppressor in a number of cancers (5-7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: PACT (protein activator of protein kinase R) is a double stranded RNA binding protein and a cellular activator of PKR (protein kinase R), a kinase that mediates the antiviral and antiproliferative actions of interferon (1). Stress signals such as serum starvation and arsenite treatment induces PACT to heterodimerize with PKR, resulting in the phosphorylation of eIF-2α and inhibition of protein synthesis (1,2). PACT has also been shown to play a role in RNA-mediated gene silencing by stimulating the activity of Dicer, thereby affecting the efficiency of miRNA accumulation and siRNA gene silencing (3,4). More recently, PACT has been shown to interact with RIG-I, a sensor of viral nucleic acids, and stimulates RIG-I-induced type I interferon production (5). Researchers have found that mutations in PACT are associated with dystonia, a movement disorder where patients develop involuntary muscle contractions and postures (6-8).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: MOB1 was first identified in yeast as a protein that binds to Mps with essential roles in the completion of mitosis and the maintenance of ploidy (1). Its Drosophila and mammalian homologs, Mats and MOB1, respectively, are involved in the Hippo signaling tumor suppressor pathway, which plays a critical role in organ size regulation and which has been implicated in cancer development (2-5). There are two MOB1 proteins in humans, MOB1α and MOB1β, that are encoded by two different genes but which have greater than 95% amino acid sequence identity (6). Both forms bind to members of the nuclear Dbf2-related (NDR) kinases, such as LATS1/2 and NDR1/2, thereby stimulating kinase activity (7-9). This binding is promoted by the phosphorylation of MOB1 at several threonine residues by MST1 and/or MST2 (5,10).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: Small non-coding RNAs are important regulators of gene expression in higher eukaryotes (1,2). Several classes of small RNAs, including short interfering RNAs (siRNAs) (3), microRNAs (miRNAs) (4), and Piwi-interacting RNAs (piRNAs) (5), have been identified. MicroRNAs are about 21 nucleotides in length and have been implicated in many cellular processes such as development, differentiation, and stress response (1,2). MicroRNAs regulate gene expression by modulating mRNA translation or stability (2). MicroRNAs function together with the protein components in the complexes called micro-ribonucleoproteins (miRNPs) (2). Among the most important components in these complexes are Argonaute proteins (1,2). There are four members in the mammalian Argonaute family and only Argonaute 2 (Ago2) possesses the Slicer endonuclease activity (1,2). Argonaute proteins participate in the various steps of microRNA-mediated gene silencing, such as repression of translation and mRNA turnover (1).

$348
100 µl
This Cell Signaling Technology antibody is conjugated to biotin under optimal conditions. The biotinylated antibody is expected to exhibit the same species cross-reactivity as the unconjugated LC3B (D11) XP® Rabbit mAb #3868.
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

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 it has also been associated with a number of physiological processes including development, differentiation, neurodegenerative diseases, infection, and cancer (3). Autophagy marker Light Chain 3 (LC3) was originally identified as a subunit of microtubule-associated proteins 1A and 1B (termed MAP1LC3) (4) and subsequently found to contain similarity to the yeast protein Apg8/Aut7/Cvt5 critical for autophagy (5). Three human LC3 isoforms (LC3A, LC3B, and LC3C) undergo post-translational modifications during autophagy (6-9). Cleavage of LC3 at the carboxy terminus immediately following synthesis yields the cytosolic LC3-I form. During autophagy, LC3-I is converted to LC3-II through lipidation by a ubiquitin-like system involving Atg7 and Atg3 that allows for LC3 to become associated with autophagic vesicles (6-10). The presence of LC3 in autophagosomes and the conversion of LC3 to the lower migrating form, LC3-II, have been used as indicators of autophagy (11).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: SMARCA1 (SNF2L) is one of the two orthologs of the ISWI (imitation switch) ATPases encoded by the mammalian genome (1). The ISWI chromatin remodeling complexes were first identified in Drosophila and have been shown to remodel and alter nucleosome spacing in vitro (2). SMARCA1 is the catalytic subunit of the nucleosome remodeling factor (NURF) and CECR2-containing remodeling factor (CERF) complexes (3-5). The NURF complex plays an important role in neuronal physiology by promoting neurite outgrowth and regulation of Engrailed homeotic genes that are involved in neuronal development in the mid-hindbrain (3). NURF is also thought to be involved in the maturation of T cells from thymocytes by regulating chromatin structure and expression of genes important for T cell development (6). The largest subunit of the NURF complex, BPTF, is required for proper development of mesoderm, endoderm, and ectoderm tissue lineages, suggesting a role for SMARCA1 in the development of the germ layers in mouse embryo (7). Disruption of the CERF complex by deletion of CECR2, an interacting partner of SMARCA1, is associated with the neural tube defect exencephaly, linking the CERF complex with regulation of neurulation (4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

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

Background: The sodium-dependent phosphate transport protein 2B (NaPi-2b, SLC34A2) is a sodium dependent inorganic phosphate (Pi) transporter that regulates phosphate homeostasis in various organs, including the small intestine, lung, liver, and testis (1). In the small intestine, NaPi-2b localizes to the intestinal brush border membrane to mediate Pi reabsorption (2). In the lung, NaPi-2b is expressed in the apical membrane of type II alveolar cells and is involved in the synthesis of surfactant (3). Mutations in the corresponding SLC34A2 gene causes pulmonary alveolar microlithiasis, a rare autosomal recessive disorder characterized by the deposition of calcium phosphate microliths throughout the lungs (4). Research studies show aberrant expression of NaPi-2b in various type of cancer, including ovarian, breast, and lung cancer (5). Chromosomal rearrangements involving SLC34A2-ROS1 are seen in gastric carcinoma and non-small cell lung cancer and result in the formation of a SLC34A2-ROS1 chimeric protein that retains a constitutive kinase activity (6,7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

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

Background: The PRNP gene encodes the major prion protein (PrP, CD230), a widely-expressed glycoprotein expressed at high levels in the central nervous system (1). While the typical cellular function of PrP is not well defined, it is a putative antioxidant and a metal-binding protein that may be involved in signal transduction (2). Prion proteins can adopt different conformations; the cellular PrPc prion protein may be converted following translation into the β-sheet-rich scrapie isoform (PrPsc) responsible for several prion diseases, including bovine spongiform encephalopathy and human Creutzfeldt-Jakob disease (3). Unlike most neurodegenerative diseases, prion diseases are infectious as prions are capable of propagating by conferring an abnormally folded state onto properly folded cellular proteins (3). In addition, the cellular PrPc has may be involved in β-amyloid peptide oligomerization and synaptic toxicity (4).

$348
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometry analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Helios (D8W4X) XP® Rabbit mAb #42427.
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Flow Cytometry

Background: Helios (Ikaros family zinc finger 2, IZKF2) is an Ikaros family transcription factor composed of several zinc fingers that mediate DNA binding and homodimerization or heterodimerization with other Ikaros family proteins (1,2). In the hematopoietic system, Helios expression is restricted to T cells and early hematopoietic progenitors (1,2). In regulatory T cells, expression of Helios contributes to an anergic phenotype by binding to the IL-2 promoter and suppressing IL-2 transcription (3). In addition, alteration of the corresponding Helios gene IZKF2 is one hallmark of low-hypodiploid acute lymphoblastic leukemia (4).

$269
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

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

Background: Zinc finger and BTB domain-containing protein 7A (LRF, Pokemon, FBI1) is a transcriptional repressor encoded by the ZBTB7A gene that belongs to the POK (POZ and Kruppel)/ZBTB (zinc finger and BTB) family (1). LRF is broadly expressed with elevated expression in a variety of cancers relative to normal tissues, including non-small cell lung cancer, breast cancer, ovarian cancer, prostate cancer, and hepatocellular carcinoma (1-8). Research studies suggest that LRF acts as an oncogene through various mechanisms including repression of the tumor suppressors ARF and Rb, and repression of the cell cycle arrest factor p21Cip1 (9-11). The LRF transcription factor plays key roles during several stages of hematopoiesis including promoting lymphoid progenitor cells to commit to B cell differentiation by repressing T cell-promoting Notch signals, and promoting cell survival during terminal erythroid differentiation through suppression of the proapoptotic factor Bim (12,13).

$489
96 assays
1 Kit
CST's PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-Histone H3 (Ser10) protein. A Histone H3 Antibody has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-Histone H3 proteins are captured by the coated antibody. Following extensive washing, a Biotinylated Phospho-Histone H3 (Ser10) Antibody is added to detect the captured phospho-Histone H3 (Ser10) protein. HRP-linked Streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of Phospho-Histone H3 (Ser10) protein.Antibodies in kit are custom formulations specific to kit.
REACTIVITY
Human, Mouse

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).

Each control slide contains formalin fixed, paraffin-embedded cell pellets: Raw 264.7 (mPD-L1 negative) and mouse bone marrow-derived macrophages (mPD-L1 positive), which serve as controls for mPD-L1 immunostaining.

Background: Programmed cell death 1 ligand 1 (PD-L1, B7-H1, CD274) is a member of the B7 family of cell surface ligands that regulate T cell activation and immune responses. The PD-L1 ligand binds the PD-1 transmembrane receptor and inhibits T cell activation. PD-L1 was discovered following a search for novel B7 protein homologs and was later shown to be expressed by antigen presenting cells, activated T cells, and tissues including placenta, heart, and lung (1-3). Similar in structure to related B7 family members, PD-L1 protein contains extracellular IgV and IgC domains and a short, cytoplasmic region. Research studies demonstrate that PD-L1 is expressed in several tumor types, including melanoma, ovary, colon, lung, breast, and renal cell carcinomas (4-6). Expression of PD-L1 in cancer is associated with tumor infiltrating lymphocytes, which mediate PD-L1 expression through the release of interferon gamma (7). Additional research links PD-L1 expression to cancers associated with viral infections (8,9).

$303
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: The ezrin, radixin, and moesin (ERM) proteins function as linkers between the plasma membrane and the actin cytoskeleton and are involved in cell adhesion, membrane ruffling, and microvilli formation (1). ERM proteins undergo intra or intermolecular interaction between their amino- and carboxy-terminal domains, existing as inactive cytosolic monomers or dimers (2). Phosphorylation at a carboxy-terminal threonine residue (Thr567 of ezrin, Thr564 of radixin, Thr558 of moesin) disrupts the amino- and carboxy-terminal association and may play a key role in regulating ERM protein conformation and function (3,4). Phosphorylation at Thr567 of ezrin is required for cytoskeletal rearrangements and oncogene-induced transformation (5). Ezrin is also phosphorylated at tyrosine residues upon growth factor stimulation. Phosphorylation of Tyr353 of ezrin transmits a survival signal during epithelial differentiation (6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: The family of Trk receptor tyrosine kinases consists of TrkA, TrkB, and TrkC. While the sequence of these family members is highly conserved, they are activated by different neurotrophins: TrkA by NGF, TrkB by BDNF or NT4, and TrkC by NT3 (1). Neurotrophin signaling through these receptors regulates a number of physiological processes, such as cell survival, proliferation, neural development, and axon and dendrite growth and patterning (1). In the adult nervous system, the Trk receptors regulate synaptic strength and plasticity. TrkA regulates proliferation and is important for development and maturation of the nervous system (2). Phosphorylation at Tyr490 is required for Shc association and activation of the Ras-MAP kinase cascade (3,4). Residues Tyr674/675 lie within the catalytic domain, and phosphorylation at these sites reflects TrkA kinase activity (3-6). Point mutations, deletions, and chromosomal rearrangements (chimeras) cause ligand-independent receptor dimerization and activation of TrkA (7-10). TrkA is activated in many malignancies including breast, ovarian, prostate, and thyroid carcinomas (8-13). Research studies suggest that expression of TrkA in neuroblastomas may be a good prognostic marker as TrkA signals growth arrest and differentiation of cells originating from the neural crest (10).

$122
20 µl
$293
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

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

Background: Caldesmon-1 is an actin filament stabilizing protein involved in the regulation of cell contraction. Binding of caldesmon-1 to actin is weakened by phosphorylation and by calmodulin in the presence of calcium. Caldesmon-1 is encoded by a single gene, which is spliced to generate a widely distributed low molecular weight form and a smooth muscle specific high molecular weight form (1,2). Caldesmon-1 is phosphorylated by the cyclin dependent kinase cdc2 and Erk1/2 MAP kinase, both of which prevent the activity of caldesmon-1 (3-5). Phosphorylation of caldesmon-1 by cdc2 is required for passage of cells through mitosis (6). Phosphorylation by Erk1/2 is important in regulating smooth muscle contraction (7). Caldesmon-1 activity may play a role in the formation of podosomes, adhesion complexes associated with the secretion of matrix metalloproteases, invasion, and metastasis (reviewed in 5).

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

Application Methods: Immunoprecipitation, Western Blotting

Background: The process of SUMO conjugation to target proteins is similar to the molecular chain of events observed with ubiquitin (1). SUMO is conjugated to target proteins through the coordinated action of the cellular SUMO conjugation machinery consisting of E1, E2, and E3 enzymes (2). The canonical SUMO E1 activating enzyme is a heterodimer consisting of SAE1 (AOS1) and UBA2 (SAE2) subunits. Mature SUMO is activated by E1 in an ATP-dependent reaction that generates adenylated SUMO, which functions as a high-energy intermediate in the formation of a thioester linkage between SUMO and Cys173 of UBA2 (3,4). SUMO is subsequently transferred from UBA2 to the SUMO E2 conjugating enzyme, UBC9 (5). Recent evidence suggests that redox regulation of UBA2 serves as a physiologic mechanism to modulate the cellular level of sumoylated target proteins (6).

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

Application Methods: Immunoprecipitation, Western Blotting

Background: SMG-1 is a member of the phosphoinositide 3-kinase-related kinase (PIKK) family, which includes ATM, ATR, mTOR, DNA-PKcs, and TRRAP (1,2). Activated by DNA damage, SMG-1 has been shown to phosphorylate p53 and hUpf1 (SMG-2) (1-4). hUpf1 is a subunit of the surveillance complex that allows degradation of messenger RNA species containing premature termination codons (PTCs). This process, known as nonsense-mediated mRNA decay (NMD), prevents the translation of truncated forms of proteins that may result in gain of function or dominant negative species. NMD occurs under normal cellular conditions as well as in response to damage (5,6). SMG-1 has also been shown to affect cell death receptor signaling and to protect cells from extrinsically induced apoptotic cell death (7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Importins belong to the karyopherin family of nuclear transport proteins (1) and are divided into two subgroups: importin α and importin β. Importins mainly function in nuclear protein import and export (2,3). Importin β1 (also known as karyopherin β1, Kpnβ1, Kpnb1, or p97) plays a key role in the nuclear import process (1-3). Nuclear import via importin β1 association with adaptor importin α (also known as karyopherin α, or Kpnα) is an essential component of the classical nuclear localization signal (NLS) pathway (4). Importin α directly recognizes the NLS present in the cargo target, prompting complex formation with importin β1. The cargo:importin α:importin β1 complex is transported across the nuclear pore complex (NPC) into the nucleus, where it is dissociated by the binding of RanGTP (1-4). Nuclear import directly via importin β1 can also occur by importin β1 recognition of the cargo protein, bypassing importin α involvement. In both cases, the importin β1/target protein interaction is mediated through the binding of importin β1 HEAT repeats with the target protein sequences (either the cargo protein itself or importin α) (5).

$303
100 µl
APPLICATIONS
REACTIVITY
Mouse

Application Methods: Western Blotting

Background: TBC1D1 is a paralog of AS160 (1) and both proteins share about 50% identity (2). TBC1D1 was shown to be a candidate gene for severe obesity (3). It plays a role in Glut4 translocation through its GAP activity (2,4). Studies indicate that TBC1D1 is highly expressed in skeletal muscle (1). Insulin, AICAR, and contraction directly regulate TBC1D1 phosphorylation in this tissue (1). Three AMPK phosphorylation sites (Ser231, Ser660, and Ser700) and one Akt phosphorylation site (Thr590) were identified in skeletal muscle (5). Muscle contraction or AICAR treatment increases phosphorylation on Ser231, Ser660, and Ser700 but not on Thr590; insulin increases phosphorylation on Thr590 only (5).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunoprecipitation, Western Blotting

Background: ASF1 was first identified in S. cerevisiae based on its ability to de-repress transcriptional silencing when overexpressed (1). While only one gene exists in yeast and Drosophila, mammalian cells contain the two highly homologous ASF1A and ASF1B genes (2). ASF1A and ASF1B function as histone chaperones, delivering histone H3/H4 dimers to CAF-1 or HIRA histone deposition complexes to facilitate replication-coupled and replication-independent nucleosome assembly on DNA (2-5). Both ASF1A and ASF1B bind to CAF-1, but only ASF1A binds to HIRA (5). In addition to playing a role in DNA replication and gene silencing, ASF1 functions in DNA damage repair, genome stability and cellular senescence. Deletion of ASF1 in yeast and Drosophila confers sensitivity to various DNA damaging agents and inhibitors of DNA replication, increases genomic instability and sister chromatid exchange, and activates the DNA damage checkpoint (6-8). Depletion of both ASF1A and ASF1B in mammalian cells results in the accumulation of cells in S phase, increased phosphorylation of H2A.X, centrosome amplification and apoptosis (9,10). ASF1A is required for the formation of senescence-associated heterochromatin foci (SAHF), with overexpression of ASF1A inducing senescence in primary cells (4). Both ASF1A and ASF1B are phosphorylated in S phase by the Tousled-like kinases TLK1 and TLK2, and are dephosphorylated when TLK1 and TLK2 are inactivated by Chk1 kinase in response to replicative stress (11,12). The function of ASF1 phosphorylation is not yet understood.