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Product listing: DeltaN p63 (E6Q3O) Rabbit mAb, UniProt ID Q9H3D4-2 #67825 to CD16 (D5Q8N) Rabbit mAb, UniProt ID P08637 #80006

$260
100 µl
APPLICATIONS
REACTIVITY
Human

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

Background: The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (1). In addition to p53, mammalian cells contain two p53 family members, p63 and p73, which are similar to p53 in both structure and function (2). While p63 can induce p53-responsive genes and apoptosis, mutation of p63 rarely results in tumors (2). Research investigators frequently observe amplification of the p63 gene in squamous cell carcinomas of the lung, head and neck (2,3). The p63 gene contains an alternative transcription initiation site that yields a truncated ΔNp63 lacking the transactivation domain, and alternative splicing at the carboxy-terminus yields the α, β, and γ isoforms (3,4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey, Rat

Application Methods: Western Blotting

Background: L-asparaginase (ASRGL1) catalyzes the conversion of L-asparagine to L-aspartate. Research studies have shown that intracellular asparagine can suppress apoptosis in a large number of human tumors (1). In addition, acute lymphocytic leukemia cells frequently depend upon serum asparagine for their viability, as they lack asparagine synthetase (ASNS). Deprivation of asparagine by L-asparaginase has therefore been developed as a therapeutic treatment for acute lymphocytic leukemia (2-3). In KRAS mutant non-small cell lung carcinoma (NSCLC) cells, PI3K/Akt signaling was shown to be required for ASNS expression, suggesting combinatorial Akt inhibition and L-asparaginase treatment as a therapeutic strategy for NSCLC (3). Research studies on a breast cancer model have furthermore shown that restriction of asparagine can suppress cancer metastasis (4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: FAM134B (family with sequence similarity 134, member B), also referred to as JK-1 and RETREG1, is a cis-Golgi endoplasmic reticulum (ER) transmembrane protein that plays a role in ER homeostasis and may contribute to several human diseases (1). FAM134B contains a conserved LC3 interacting domain (LIR) that facilitates binding to LC3 and GABARAP family members and targets impaired ER to the autophagsome for degradation by ER-phagy (2). Deletion of FAM134B leads to ER expansion and stress-induced apoptosis (2). Expression of FAM134B has been linked to a number of pathological conditions, including viral infection, cancer, and neuronal disorders (1). FAM134B can potentially inhibit viral infection, as demonstrated by studies of FAM134B knockouts that resulted in significantly higher rates of Ebola virus replication (3). Mutations in FAM134B that lead to an accumulation of mis-folded proteins have also been associated with neuronal sensory disorders (2, 4, 5). The expression and mutational state of FAM134B can also have varying effects on cancer. Oncogenic effects of FAM134B were described in esophageal squamous carcinoma; whereas, it appears to have tumor suppressor activity in colorectal cancer (6-8).

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

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

Background: Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: Carbohydrate-responsive element-binding protein (ChREBP) is a glucose-responsive transcription factor that regulates glycolytic and lipogenic gene expression (1). High levels of glucose induce the transcriptional activity of ChREBP. ChREBP is most abundant in tissues of de novo lipogenesis and forms a heterotetramer with its binding partner MLX to bind to the promoter regions of its target genes (2). ChREBP regulates fatty acid synthesis, glucose homeostasis and insulin sensitivity in white adipose tissue. In white adipose tissue, ChREBP isoform α (ChREBP-α) activates the expression of the potent ChREBP isoform β (ChREBP-β) in a glucose-dependent manner. ChREBP-β expression levels predict insulin sensitivity in human white adipose tissue (1). In addition, research studies have shown that Akt2 induces the transcriptional activity of ChREBP-β to stimulate de novo lipogenesis in brown adipose tissue for fuel storage (3). Furthermore, ChREBP-β is a potent activator of lipogenesis in the liver (4).

$260
100 µl

Background: N6-methyladenosine (m6A) is a post-transcriptional modification found in various RNA subtypes. While the presence of m6A in RNA was described decades ago, the lack of tools has made interrogating the epitranscriptomic landscape challenging (1,2). With the emergence of new technologies such as miCLIP and NG-RNA-seq, researchers have been able to show that m6A is a biologically relevant mark in mRNA that is enriched in 3’ UTRs and stop codons (3,4). The m6A writer complex consists of a core heterodimer of methyltransferase-like protein 3 (METTL3) and methytransferase-like protein 14 (METTL14), and the additional regulatory proteins Virlizer/VIRMA and Wilms tumor 1-associated protein (WTAP) (5). METTL3 is the catalytic methyltransferase subunit and METTL14 is the target recognition subunit that binds to RNA (6). The Virilzer/VIRMA protein directs m6A methylation to the 3’ UTRs and stop codons, and WTAP targets the complex to nuclear speckles, which are sites of RNA processing (7). Less is known about readers and erasers of m6A, and while the fat mass and obesity-associated protein FTO was the first discovered m6A demethylase, subsequent studies demonstrated that this enzyme may prefer the closely related m6Am mark in vivo (8,9). ALKBH5 was later shown to be a bona fide m6A demethylase enzyme, contributing to the idea that the m6A modification is dynamically regulated (10). Readers of the m6A mark include the YTH protein family, which can bind to m6A and influence mRNA stability and translation efficiency (3,11-13). The m6A mark and machinery have been shown to regulate a variety of cellular functions, including RNA splicing, translational control, pluripotency and cell fate determination, neuronal function, and disease (1, 14-17). The m6A writer complex has been linked to various cancer types including AML and endometrial cancers (18,19). Additionally, m6A has been implicated in resistance to chemotherapy (20).

$129
20 µl
$303
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: IHC-Leica® Bond™, Immunohistochemistry (Paraffin), Immunoprecipitation, Western Blotting

Background: Semaphorin-4D/CD100 (Sema4D) is a disulfide-linked homodimeric type 1 transmembrane glycoprotein belonging to the class IV family of membrane bound semaphorins. The extracellular domain of Sema4D contains a cysteine-rich semaphorin-like domain, an Ig-like domain, and a PSI domain (1). Research studies have suggested that the cytoplasmic domain has a signaling function as it is phosphorylated on serine residues (2). Initial studies involving Sema4D revealed that it was implicated in axon guidance within the central nervous system through binding its high affinity receptor, plexin-B1 (3). Sema4D function has also been extensively characterized in the immune system and is the first semaphorin found to be expressed on the surface of many types of immune cells (4-6). In the immune system, CD72 serves as a low-affinity receptor for Sema4D (7) and research studies have shown that Sema4D not only regulates T-cell activation (8,9) but is also involved in the regulation of B-cell survival and differentiation (10). Many of the physiologic effects of Sema4D in the immune system are regulated by a soluble extracellular domain-containing fragment generated through proteolytic cleavage (11).Sema4D has also been implicated in oncogenesis as research studies have demonstrated overexpression in multiple types of solid tumors (12,13). The role of Sema4D in oncogenesis, in part, has been linked to its ability to promote tumor angiogenesis (14), cell invasion (15), and immunosuppression through enhancement of myeloid derived suppressor cell function (16).

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

Application Methods: Western Blotting

Background: Malic enzymes catalyze oxidative decarboxylation of malate to pyruvate (1). The malic enzyme family in mammalian cells includes the cytosolic malic enzyme 1 (ME1) and two mitochondrial malic enzymes (ME2 and ME3) (1, 2). ME1 and ME2 are critical for tumor cell growth and their expression is repressed by tumor suppressor p53 (2). Reduced expression of ME1 and ME2 reciprocally increases the levels and activation of p53, promoting p53-mediated senescence (2). Research studies show ME3 is essential for the survival of pancreatic ductal adenocarcinoma following genomic deletion of ME2 (3). Deletion of ME3 is lethal to ME2-null cancer cells, which has been suggested to provide a potential therapeutic opportunity using collateral lethality (3, 4).

$303
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

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

Background: Tau is a heterogeneous microtubule-associated protein that promotes and stabilizes microtubule assembly, especially in axons. Six isoforms with different amino-terminal inserts and different numbers of tandem repeats near the carboxy terminus have been identified, and tau is hyperphosphorylated at approximately 25 sites by Erk, GSK-3, and CDK5 (1,2). Phosphorylation decreases the ability of tau to bind to microtubules. Neurofibrillary tangles are a major hallmark of Alzheimer's disease; these tangles are bundles of paired helical filaments composed of hyperphosphorylated tau. In particular, phosphorylation at Ser396 by GSK-3 or CDK5 destabilizes microtubules. Furthermore, research studies have shown that inclusions of tau are found in a number of other neurodegenerative diseases, collectively known as tauopathies (1,3).

$303
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: LAT, a transmembrane adaptor protein expressed in T, NK and mast cells, is an important mediator for T cell receptor (TCR) signaling (1). Upon TCR engagement, activated Zap-70 phosphorylates LAT at multiple conserved tyrosine residues within SH2 binding motifs, exposing these motifs as the docking sites for downstream signaling targets (2,3). The phosphorylation of LAT at Tyr171 and Tyr191 enables the binding of Grb2, Gads/SLP-76, PLCγ1 and PI3 kinase through their SH2 domain and translocates them to the membrane. This process eventually leads to activation of the corresponding signaling pathways (1-4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: Neuronal Cell Adhesion Molecule, or NRCAM, belongs to the immunoglobulins Cell Adhesion Molecules (CAM's) superfamily (1). NRCAM, an ankyrin-binding protein, contributes to the neurite outgrowth by providing directional signaling during axonal cone growth (2, 3, 4). Additionally, it plays a role in mediating the interaction between axons and Schwann cells and contributes to the formation and maintenance of Nodes of Ranvier (5, 6, 7, 8). NRCAM also plays an important role in the establishment of dendritic spines in developing cortical neurons (9). NRCAM is expressed in non-neuronal cells, mostly in endothelial cells (10).

$348
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct immunofluorescent analysis. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated β-Amyloid (D54D2) XP® Rabbit mAb #8243.
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunofluorescence (Frozen)

Background: Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms (1). The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage (1). The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques in Alzheimer's disease (1). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion of this protein (2-5). Phosphorylation at Thr668 (a position corresponding to the APP695 isoform) by cyclin-dependent kinase is cell-cycle dependent and peaks during G2/M phase (4). APP phosphorylated at Thr668 exists in adult rat brain and correlates with cultured neuronal differentiation (5,6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: Cancer/testis antigens (CTAs) are a family of more than 100 proteins whose normal expression is largely restricted to immune privileged germ cells of the testis, ovary, and trophoblast cells of the placenta. Although most normal somatic tissues are void of CTA expression, due to epigenetic silencing of gene expression, their expression is upregulated in a wide variety of human solid and liquid tumors (1,2). As such, CTAs have garnered much attention as attractive targets for a variety of immunotherapy-based approaches to selectively attack tumors (3).

C18 Reversed-phase solid phase extraction columns are useful tools for purifying and desalting peptides in preparation for PTMScan protocols and other mass spectrometry applications.
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

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

Background: The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Major types of intermediate filaments are specifically expressed in particular cell types: cytokeratins in epithelial cells, glial fibrillary acidic protein (GFAP) in glial cells, desmin in skeletal, visceral, and certain vascular smooth muscle cells, vimentin in cells of mesenchymal origin, and neurofilaments in neurons. GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape (1). In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes (2). In addition, GFAP intermediate filaments are also present in nonmyelin-forming Schwann cells in the peripheral nervous system (3).

$108
250 PCR reactions
500 µl
SimpleChIP® Human HUWE1 Intron 26 Control Primers contain a mix of forward and reverse PCR primers that are specific to intron 26 of the human HECT, UBA and WWE domain containing 1, E3 ubiquitin protein ligase gene. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitations (ChIP). Primers have been optimized for use with SimpleChIP® Universal qPCR Master Mix #88989 and have been tested in conjunction with SimpleChIP® Plus Enzymatic Chromatin IP Kits #9004 and #9005 and ChIP-validated antibodies from Cell Signaling Technology®.
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, Monkey, Mouse

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

Background: The polycomb group (PcG) proteins contribute to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). CBX4 is a component of the PRC1 complex, which together with Ring1 strongly enhances the E3 ubiquitin ligase activity of the Ring2 catalytic subunit (7,8). CBX4 itself is a SUMO E3 ligase, and its function influences EMT, DNA damage response, tumor angiogenesis, and self-renewal (9-13).

$269
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: IHC-Leica® Bond™, Immunohistochemistry (Paraffin), Western Blotting

Background: Macrophage-colony stimulating factor (M-CSF, CSF-1) receptor is an integral membrane tyrosine kinase encoded by the c-fms proto-oncogene. M-CSF receptor is expressed in monocytes (macrophages and their progenitors) and drives growth and development of this blood cell lineage. (1-3). Binding of M-CSF to its receptor induces receptor dimerization, activation, and autophosphorylation of cytoplasmic tyrosine residues used as docking sites for SH2-containing signaling proteins (4). There are at least five major tyrosine autophosphorylation sites. Tyr723 (Tyr721 in mouse) is located in the kinase insert (KI) region. Phosphorylated Tyr723 binds the p85 subunit of PI3 kinase as well as PLCγ2 (5). Phosphorylation of Tyr809 provides a docking site for Shc (5). Overactivation of this receptor can lead to a malignant phenotype in various cell systems (6). The activated M-CSF receptor has been shown to be a predictor of poor outcome in advanced epithelial ovarian carcinoma (7) and breast cancer (8).

$499
96 assays
1 Kit
The FastScan™ Total Zap-70 ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Zap-70. 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 Zap-70 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 Zap-70. Antibodies in kit are custom formulations specific to kit.
REACTIVITY
Human, Mouse

Background: The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Calretinin (29 kDa calbindin, calbindin 2) is a calcium-binding protein of the EF-hand family encoded by the CALB2 gene. It is differentially expressed from homologous family member calbindin-d28k in distinct neuronal populations of the retina, auditory system, and cerebellar granule cells (1, 2), and acts as a marker for specific neuronal subsets of the subthalamic nucleus and the substantia nigra (3). Calretinin has been shown to play an important role in modulating neuronal excitability and the induction of long-term potentiation (1). Research has shown that, pathologically, calretinin is a selective marker for epithelial mesothelioma, making it a diagnostic tool to differentiate from adenocarcinomas (4).

$59
125 ml
This product is a ready-to-use buffer, optimized for the dilution and incubation of both conjugated and unconjugated antibodies in flow cytometry assays (F). The buffer is formulated with 1X phosphate-buffered saline (PBS) compatible for use with live or fixed cells, and contains bovine serum albumin (BSA) to prevent antibody aggregation. Cell Signaling Technology recommends using this buffer according to our protocols for flow cytometry-approved antibodies to ensure accurate and reproducible results.
APPLICATIONS

Application Methods: Flow Cytometry

Premium Methanol from Cell Signaling Technology is submicron filtered and 99.9% pure. It is recommended for use as a fixative or permeabilization agent in immunofluorescence and flow cytometry assays.Important: Not all CST primary antibodies require samples to be prepared with methanol fixation or permeabilization prior to immunostaining. Please refer to the antibody datasheet to determine if methanol is recommended.
$108
250 PCR reactions
500 µl
SimpleChIP® Human FOXN3 Intron3 Primers contain a mix of forward and reverse PCR primers that are specific to intron 3 of the human forkhead box N3 (FOXN3) gene. These primers can be used to amplify DNA that has been isolated using chromatin immunoprecipitation (ChIP). Primers have been optimized for use in quantitative real-time PCR and have been tested in conjunction with SimpleChIP® Enzymatic Chromatin IP Kits #9004 and #9005, ChIP-validated antibodies from Cell Signaling Technology®, and SimpleChIP® Universal qPCR Master Mix #88989.
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

Application Methods: Immunoprecipitation, Western Blotting

Background: Peroxisome proliferator-activated receptor γ (PPARγ) is a member of the ligand-activated nuclear receptor superfamily and functions as a transcriptional activator (1). PPARγ is preferentially expressed in adipocytes as well as in vascular smooth muscle cells and macrophage (2). Besides its role in mediating adipogenesis and lipid metabolism (2), PPARγ also modulates insulin sensitivity, cell proliferation and inflammation (3). PPARγ transcriptional activity is inhibited by MAP kinase phosphorylation of PPARγ at Ser84 (4,5).

$129
20 µl
$303
100 µl
APPLICATIONS
REACTIVITY
Mouse, Rat

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

Background: Adenosine Receptor A2a (A2AR) is a G-protein-coupled receptor (GPCR). As a member of the purinergic adenosine receptors (A1, A2, and A3), A2AR activates classic G-protein signaling pathways upon binding of adenosine (1). Adenosine is present in all cells and extracellular fluids. Adenosine signaling, via A2AR, is mobilized during both physiological and pathological conditions. For example, adenosine, via A2AR, modulates neuronal function, acting to fine-tune neuronal function (2). A2AR function is modulated, in part, by its ability to form functional heteromers with other GPCRs, including dopamine receptors (D1 and D3), metabotropic glutamate receptors (mGluR5), and others (3). In the brain, A2AR is enriched in the basal ganglia, suggesting that A2AR may be a potential drug target for neurodegenerative diseases like Parkinson’s disease, drug addiction, and psychiatric disorders (4). Outside of the brain, A2AR may act as an immune checkpoint molecule to maintain an immunosuppressive tumor microenvironment, an environment that exhibits relatively elevated adenosine levels (5, 6).

$269
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: IHC-Leica® Bond™, Immunohistochemistry (Paraffin)

Background: Chromosomal translocations result in misregulation of the proto-oncogene BCL6 in patients with B cell-derived non-Hodgkin's lymphoma (1). The BCL6 gene is selectively expressed in mature B cells and encodes a nuclear phosphoprotein that belongs to the BTB/POZ zinc finger family of transcription factors (2,3). BCL6 protein can bind to target DNA sequences of Stat6 and, analogous to Stat6, modulate the expression of interleukin-4-induced genes (4). Furthermore, BCL6 restrains p53-dependent senescence, making BCL6-active tumors functionally p53-negative (5). The mitogen-activated protein kinases, Erk1 and Erk2, but not JNK, phosphorylate BCL6 at multiple sites. Phosphorylation of BCL6 at Ser333 and Ser343 results in degradation of BCL6 by the ubiquitin/proteasome pathway in B cells (6,7). In addition, BCL6 is acetylated and its transcriptional repressor function is inhibited by the transcriptional co-activator p300 (8).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: MIF (macrophage migration inhibitory factor) is a pleiotropic cytokine that stimulates pro-inflammatory, chemotactic, and growth responses in cells (1). MIF binds its cognate receptor (a CD74/CD44 complex) to activate multiple signaling pathways such as Src, ERK, MAPK, Akt, and suppress p53-induced apoptosis (2). The interaction of MIF with non-cognate chemokine receptors CXCR2 and CXCR4 promotes chemotaxis that enables recruitment of monocytes/neutrophils and T cells (3). During an innate immune response, MIF has been shown to repress the inhibitory effects of glucocorticoids on macrophages and T cells, thus promoting host inflammation and immunity (4). MIF may also play roles in the progression of other disease processes, including cancer cell proliferation and metastasis, and angiogenesis (5), atherosclerotic plaque formation following myocardial ischemia (6), and autoimmune pathogenesis (7). MIF has thus been proposed as a promising therapeutic drug target for multiple indications.

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

Application Methods: Western Blotting

Background: Quaking (QKI) is a member of the signal transduction and activator of RNA (STAR) protein family of RNA binding proteins (1,2). Mutations in the mouse Qki locus results in impaired myelin formation resulting in tremors (3). QKI proteins exist as homodimers, and disruption of the dimerization process is lethal in mice (2). QKI exists in different isoforms that differ in their C-terminus, resulting in unique subcellular localizations (4). The nuclear isoform of QKI, QKI-5, is involved in regulation of alternative splicing of MAG mRNA, which encodes for a protein important for myelin sheath formation and maintenance (4,5). QKI has also been implicated in schizophrenia and oligodendrocyte differentiation (6). QKI can function as a tumor suppressor, as it is regulated by p53 to stabilize miRNAs that regulate TGF-Β signaling (7).

$305
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated p16 INK4A (D7C1M) Rabbit mAb #80772.
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry

Background: Members of the INK4 family of cyclin dependent kinase inhibitors include p16INK4A, p15INK4B, p18INK4C and p19INK4D. The INK4 family members inhibit cyclin dependent kinases 4 and 6 (CDK4 and CDK6), causing cell cycle arrest in G1 phase. The INK4A-ARF-INK4B locus on chromosome 9p21, frequently lost in human cancer, encodes the INK4 family members p16INK5A and p15INK4B, as well as the unrelated protein, ARF (1).p16 INK4A expression, typically repressed in the absence of stress, is thought to drive cells into senescence, and p16 INK4A expression is a commonly used marker of senescent cells (2). p16INK4A protein expression is often altered in human cancer (3,4), and high expression is currently used as a predictive biomarker in cervical cancer (5).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: CD64 (FcgammaRI), CD32 (FcgammaRII) and CD16 (FcgammaRIII) are three classes of the immunoglobulin superfamily. CD64 has a high affinity for IgG with three Ig-like domains while CD32 and CD16 have low affinities with two Ig-like domains. Two genes encode CD16-A and CD16-B resulting only in a 6 amino acid difference in their ectodomains. However, CD16-A has a transmembrane anchor versus CD16-B, which has a glycosylphosphatidylinositol (1). CD64, CD32 and CD16 are membrane glycoproteins that are expressed by all immunologically active cells and trigger various immune functions (activate B cells, phagocytosis, antibody-dependent cellular cytotoxicity, immune complex clearance and enhancement of antigen presentation) (2). CD16 cross-linking induces tyrosine phosphorylation (Tyr394) of Lck in NK cells (3). CD32 has tyrosine-based activation motifs in the cytoplasmic domain in contrast to CD16, which associates with molecules possessing these motifs (1).