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Product listing: ATM (11G12) Mouse mAb, UniProt ID Q13315 #92356 to Phospho-Chk1 (Ser296) (D3O9F) Rabbit mAb, UniProt ID O14757 #90178

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Ataxia telangiectasia mutated kinase (ATM) is a serine/threonine kinase that regulates cell cycle checkpoints and DNA repair (1). Activation of ATM by autophosphorylation on Ser1981 occurs in response to exposed DNA double stranded breaks. ATM kinase regulates a number of proteins involved in cell cycle checkpoint control, apoptosis, and DNA repair. Known substrates include p53, Chk2, Chk1, CtIP, 4E-BP1, BRCA1, RPA3, H2A.X, SMC1, FANCD2, Rad17, Artemis, Nbs1, and the I-2 regulatory subunit of PP1 (1,2). Mutations in the corresponding ATM gene result in ataxia telangiectasia (AT), an autosomal recessive disease characterized by uncoordinated muscle movement and neurodegeneration. Cells from AT patients display defective DNA damage-induced checkpoint activation, sensitivity to radiation, and a higher frequency of chromosome breakage (3,4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

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

Background: TNF receptor-associated protein 1 (TRAP1), also known as HSP75, is a mitochondrial chaperone and ATPase that was originally identified as a protein that interacts with the TNF receptor. Although a member of the HSP90 family, TRAP1 is not heat-inducible but is upregulated by glucose deprivation, oxidative injury, and UV irradiation. An amino-terminal mitochondrial localization sequence results in localization of TRAP1 within mitochondria (1). Overexpression of TRAP1 decreases oxidative stress, suggesting a protective role in ischemia injury (2). Research studies demonstrate that silencing of TRAP1 enhances cytochrome C release and apoptosis, with additional evidence indicating that TRAP1 can protect cells from cell death by inhibiting the generation of reactive oxygen species (3). TRAP1 is a substrate of the mitochondrial serine/threonine kinase PINK1, whose corresponding gene is mutated in some forms of early-onset Parkinson's disease (PD). PINK1 protects cells from oxidative stress-induced cell death by suppressing release of cytochrome C from mitochondria. PD-linked PINK1 mutations impair the ability of PINK1 to phosphorylate TRAP1 and leads to impaired cell survival (4). Finally, TRAP1 alleviates α-synuclein induced toxicity and rescues the PINK1 loss-of-function phenotype (5).

$303
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse

Application Methods: Immunoprecipitation, Western Blotting

Background: Embryonic stem cells (ESC) derived from the inner cell mass of the blastocyst are unique in their pluripotent capacity and potential for self-renewal (1). Research studies demonstrate that a set of transcription factors that includes Oct-4, Sox2, and Nanog forms a transcriptional network that maintains cells in a pluripotent state (2,3). Chromatin immunoprecipitation experiments show that Sox2 and Oct-4 bind to thousands of gene regulatory sites, many of which regulate cell pluripotency and early embryonic development (4,5). siRNA knockdown of either Sox2 or Oct-4 results in loss of pluripotency (6). Induced overexpression of Oct-4 and Sox2, along with additional transcription factors Klf4 and c-Myc, can reprogram both mouse and human somatic cells to a pluripotent state (7,8). Additional evidence demonstrates that Sox2 is also present in adult multipotent progenitors that give rise to some adult epithelial tissues, including several glands, the glandular stomach, testes, and cervix. Sox2 is thought to regulate target gene expression important for survival and regeneration of these tissues (9).

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

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

Background: Wolf-Hirschhorn syndrome candidate 1-like protein 1 (WHSC1L1), also known as histone-lysine N-methyltransferase NSD3, is a SET domain-containing histone methyltransferase protein that methylates histone H3 on lysine 4 (H3K4me) and lysine 27 (H3K27me) (1). Methylation of histone H3 lysine 4 is associated with transcriptional activation, while methylation of histone H3 lysine 27 is associated with transcriptional repression. WHSC1L1 can function as an oncogene or a tumor suppressor protein, depending on cell context, and has been shown to regulate expression of a number of genes associated with cell cycle (2). Amplification and/or increased expression of WHSC1L1 in breast, lung, and liver cancer increases growth and survival, and is associated with poor prognosis (2-5). In addition, the NSD3 gene has been found fused with the nuclear pore complex protein NUP98 gene in acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL), and fused with the NUT gene in NUT midline carcinomas (6-10).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: Tripartite motif-containing protein 21 (TRIM21) is a cytosolic antiviral protein that is induced by interferons and acts as a cytosolic IgG receptor (1,2).  TRIM21 is an activator of the innate immune system and targets antibody bound virions to the proteasome via its E3 ubiquitin ligase activity, leading to degradation of the virions in the cytosol (3,4). Interestingly, TRIM21 has also been shown to negatively regulate pathogen-induced interferon production through the polyubiquitin-mediated degradation of multiple targets including IRF3, IRF7, and DDX41 (5,6,7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

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

Background: The mannose receptor (MR/CLEC13D/CD206/MMR/MRC1/Macrophage mannose receptor 1) is an endocytic receptor expressed by populations of dendritic cells, macrophages and nonvascular endothelium (1). The mannose receptor is a heavily glycosylated type I transmembrane protein with three types of extracellular domains and a short carboxy-terminal cytoplasmic domain with no apparent signaling motif (2-4). The extracellular portion of the protein is made up of a CR domain, which binds sulfated glycans, an FNII domain, which binds collagens, and eight C-type lectin domains, which bind carbohydrates containing mannose, fucose or GlcNAc (4-7). The receptor recycles between the plasma membrane and early endosomes (8). Functions include a role in antigen cross-presentation, clearance of endogenous proteins, pathogen detection and trafficking through lymphatic vessels (9-12).

$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 S100A9 (D5O6O) Rabbit mAb #72590.
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry

Background: S100A8 and S100A9 are calcium-binding proteins that form a noncovalent heterodimer present in monocytes, neutrophils, macrophages, and some epithelial cells (1, 2). S100A8 and S100A9 are secreted by a tubulin-dependent mechanism during inflammatory conditions and have antimicrobial and chemotactic functions (3-5). Extracellular S100A8/S100A9 also induces an inflammatory response in endothelial cells, including induction of proinflammatory chemokines and adhesion molecules and increased vascular permeability (6). S100A8/S100A9 induces and recruits myeloid-derived suppressor cells (MDSC) in tumor-bearing mice (7). MDSC produce additional S100A8/S100A9 themselves, resulting in a positive feedback mechanism that sustains MDSC accumulation (7). S100A8/S100A9 is also highly expressed in psoriatic skin, where it directly upregulates transcription of complement protein C3, which contributes to disease (8). In addition, tumor-infiltrating myeloid cells induce expression of S100A8 and S100A9 in cancer cells, which increases invasiveness and metastasis (9).

$489
96 assays
1 Kit
The PathScan® Phospho-p90RSK (Thr359) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p90RSK protein phosphorylated at Thr359. A phospho-p90RSK (Thr359) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-p90RSK proteins can be captured by the coated antibody. Following extensive washing, a p90RSK mouse antibody is added to detect the captured phospho-p90RSK protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of p90RSK phosphorylated at Thr359.Antibodies in this kit are custom formulations specific to kit.
REACTIVITY
Human, Monkey, Rat

Background: The 90 kDa ribosomal S6 kinases (RSK1-4) are a family of widely expressed Ser/Thr kinases characterized by two nonidentical, functional kinase domains (1) and a carboxy-terminal docking site for extracellular signal-regulated kinases (ERKs) (2). Several sites both within and outside of the RSK kinase domain, including Ser380, Thr359, Ser363, and Thr573, are important for kinase activation (3). RSK1-3 are activated via coordinated phosphorylation by MAPKs, autophosphorylation, and phosphoinositide-3-OH kinase (PI3K) in response to many growth factors, polypeptide hormones, and neurotransmitters (3).

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

Application Methods: Immunoprecipitation, Western Blotting

Background: ATP-dependent chromatin remodeling complexes play an essential role in the regulation of various nuclear processes, such as gene expression, DNA replication, and repair (1,2). The SWI/SNF chromatin remodeling complex consists of more than 10 subunits with a single molecule of the ATPase catalytic subunit BRM or BRG1, but not both. The activities of these two subunits drive the disruption of histone-DNA contacts that lead to changes in accessibility of crucial regulatory elements within chromatin (2-5). The BRM/BRG1 containing SWI/SNF complexes are recruited to target promoters by transcription factors, such as nuclear receptors, p53, RB, and BRCA1 to regulate gene activation, cell growth, the cell cycle, and differentiation processes (1,6-9).PBRM1/BAF180 is a unique member of the SWI/SNF complex PBAF, which binds to kinetochores in mitotic chromatin (10,11). PBAF is involved in nuclear receptor-mediated transcription and retinoic acid driven gene activation (12, 13). PBRM1/BAF180 has been shown to be a potent tumor suppressor, as it is the second-most mutated gene in renal carcinomas (14). Mutations of PBRM1/BAF180 have also been shown to be involved breast cancer, and low expression relates to poorer prognosis (15,16)

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

Application Methods: Chromatin IP, Chromatin IP-seq, Immunohistochemistry (Paraffin), Immunoprecipitation, Western Blotting

Background: ATP-dependent chromatin remodeling complexes play an essential role in the regulation of nuclear processes such as transcription and DNA replication and repair (1,2). The SWI/SNF chromatin remodeling complex consists of more than 10 subunits and contains a single molecule of either BRM or BRG1 as the ATPase catalytic subunit. The activity of the ATPase subunit disrupts histone-DNA contacts and changes the accessibility of crucial regulatory elements to the chromatin. The additional core and accessory subunits play a scaffolding role to maintain stability and provide surfaces for interaction with various transcription factors and chromatin (2-5). The interactions between SWI/SNF subunits and transcription factors, such as nuclear receptors, p53, Rb, BRCA1, and MyoD, facilitate recruitment of the complex to target genes for regulation of gene activation, cell growth, cell cycle, and differentiation processes (1,6-9).

$303
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Necroptosis, a regulated pathway for necrotic cell death, is triggered by a number of inflammatory signals including cytokines in the tumor necrosis factor (TNF) family, pathogen sensors such as toll-like receptors (TLRs), and ischemic injury (1,2). The process is negatively regulated by caspases and is initiated through a complex containing the RIP1 and RIP3 kinases, typically referred to as the necrosome. Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase that was identified as downstream target of RIP3 in the necroptosis pathway (3,4). During necroptosis RIP3 is phosphorylated at Ser227, which recruits MLKL and leads to its phosphorylation at Thr357 and Ser358 (3). Knockdown of MLKL through multiple mechanisms results in inhibition of necroptosis (3-5). While the precise mechanism for MLKL-induced necroptosis is unclear, some studies have shown that necroptosis leads to oligomerization of MLKL and translocation to the plasma membrane, where it effects membrane integrity (6-9).

$303
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunohistochemistry (Paraffin), Western Blotting

Background: Aurora kinases belong to a highly conserved family of mitotic serine/threonine kinases with three members identified among mammals: Aurora A, B, and C (1,2). Studies on the temporal expression pattern and subcellular localization of Aurora kinases in mitotic cells suggest an association with mitotic structure. Aurora kinase functional influences span from G2 phase to cytokinesis and may be involved in key cell cycle events such as centrosome duplication, chromosome bi-orientation and segregation, cleavage furrow positioning, and ingression (3). Aurora A is detected at the centrosomes, along mitotic spindle microtubules, and in the cytoplasm of mitotically proliferating cells. Aurora A protein levels are low during G1 and S phases and peak during the G2/M phase of the cell cycle. Phosphorylation of Aurora A at Thr288 in its catalytic domain increases kinase activity. Aurora A is involved in centrosome separation, maturation, and spindle assembly and stability. Expression of Aurora B protein also peaks during the G2/M phase of the cell cycle; Aurora B kinase activity peaks at the transition from metaphase to the end of mitosis. Aurora B associates with chromosomes during prophase prior to relocalizing to the spindle at anaphase. Aurora B regulates chromosome segregation through the control of microtubule-kinetochore attachment and cytokinesis. Expression of both Aurora A and Aurora B during the G2/M phase transition is tightly coordinated with histone H3 phosphorylation (4,5); research investigators have observed overexpression of these kinases in a variety of human cancers (2,4). Aurora C localizes to the centrosome from anaphase to cytokinesis and both mRNA and protein levels peak during G2/M phase. Although typical Aurora C expression is limited to the testis, research studies report overexpression of Aurora C is detected in various cancer cell lines (6).

PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology (CST) for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® Technology enables researchers to isolate, identify, and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity, providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur. For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/common/content.jsp?id=ptmscan-services.
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Western Blotting

Background: Heterogeneous nuclear ribonucleoprotein C1/C2 (hnRNP C1/C2) has multiple biological functions including transcriptional regulation, DNA repair, and RNA processing. hnRNP C1/C2 acts as a ‘molecular ruler’ in the mRNA processing pathway, committing nascent transcripts from the chromatin template to the mRNA export pathway once the nascent transcript becomes longer than 200-300 nucleotides (1). hnRNP C1/C2 associates with SWI/SNF and NurD family members to form the locus control region (LCR)-associated remodeling complex (LARC), which binds to β-globin gene promoter to prevent transcriptional silencing. Studies indicate that without hnRNP C1/C2, LARC does not associate with its target DNA sequence (2,3). hnRNP C1/C2 and other hnRNP family members interact with DNA damage response (DDR) proteins (4). hnRNP proteins regulate double stranded break (DSB) repair by promoting either homologous recombination (HR) or non-homologous end joining (NHEJ) (4). hnRNP C1/C2 downregulates the expression of miR-21, which leads to the increased expression of programmed cell death 4 (PDCD4) protein in glioblastoma multiforme (GBM) (5). Research studies have shown that silencing of hnRNP C1/C2 renders GBM cells more susceptible to apoptosis (5).

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: Regulation of actin dynamics at the leading edge of the cell drives cell adhesion, migration, and invasion. These processes are crucial for normal development, immune cell function, angiogenesis and cell division, and are dysregulated in diseases such as metastatic cancer (1, 2).Lamellipodin (Lpd) is a member of the Mig-10/RIAM/lamellipodin (MRL) family of adaptor proteins. Lamellipodin regulates the formation of lamellipodia at the leading edge through its interaction with and recruitment of actin-modulating proteins such as Ena/VASP (3) and the Scar/WAVE complex (4). Lamellipodin and Mena cooperate with endophilin to regulate EGFR endocytosis (5). MRL proteins can also form complexes with talin and activated integrins, guiding the formation of cell protrusions and the direction of cell migration (6).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: The pannexin family (pannexin-1, -2, and -3; PANX1-3) of gap junction proteins has homology to the invertebrate innexins and display distinct expression patterns (1). Pannexin-1 is widely expressed, with highest expression in the heart, brain, skeletal muscle, testis, and ovary (1,2). Pannexin-2 is predominately expressed in the brain (1,2) and pannexin-3 is found within the skin and connective tissues (1,3). Connexin family gap junction proteins form hemichannels that align adjacent cells, creating functional intercellular channels that are permeable to ions and small molecules. In contrast, pannexin proteins may not function as gap junction proteins since pannexins on adjacent cells may not align to form complete channels (3). These pannexin “hemichannels” may play a role in inflammation, apoptosis, and neuronal signaling by allowing permeability of ions, ATP, and potentially other small molecules into the extracellular space (4-6). Pannexin-1 can be activated by effector caspases (caspase-3 and -7), which leads to release of signal molecules that promote phagocytosis of apoptotic cells (7).

$111
20 µl
$260
100 µl
APPLICATIONS
REACTIVITY
Human

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

Background: The triggering receptor expressed on myeloid cells 2 (TREM2) protein is an innate immune receptor that is expressed on the cell surface of microglia, macrophages, osteoclasts, and immature dendritic cells (1). The TREM2 receptor is a single-pass type I membrane glycoprotein that consists of an extracellular immunoglobulin-like domain, a transmembrane domain, and a cytoplasmic tail. TREM2 interacts with the tyrosine kinase-binding protein DAP12 to form a receptor-signaling complex (2). The TREM2 protein plays a role in innate immunity and a rare functional variant (R47H) of TREM2 is associated with the late-onset risk of Alzheimer’s disease (1,3). Research studies using mouse models of Alzheimer’s disease indicate that deficiency and haploinsufficiency of TREM2 can lead to increased β-amyloid (Aβ) accumulation as a result of dysfunctional microglia response (4). These results agree with the distribution of TREM2 in human brain regions (e.g., white matter, the hippocampus, and neocortex) that are involved in Alzheimer's disease pathology (2). In addition, amyloid plaque formation induces expression of TREM2 and amyloid phagocytosis (5). Loss-of-function mutations in the corresponding TREM2 or DAP12 genes can result in Nasu-Hakola disease, a rare form of progressive presenile dementia that results from polycystic osseous lesions (6). TREM2 membrane shedding occurs by cleavage at the extracellular site between H157/S158 generating an N-terminal shedded fragment and a membrane bound C-terminal fragment (7, 8).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

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

Background: HOXB13 is a member of the HOXB cluster which, along with the HOXA, HOXC, and HOXD clusters, governs embryonic patterning along the cranio-caudal axis (1,2). HOXB13 plays a key role in the development of the ventral prostate, where it is expressed highly from the embryonic stage through adulthood (3,4). Research studies have shown that both overexpression and RNA interference can inhibit the growth of prostate cancer cells. HOXB13 can function as a tumor suppressor by negatively regulating growth through repression of TCF4 and androgen receptor (AR) signaling (4,5). However, HOXB13 has also been shown to be overexpressed in more invasive prostate cancers, breast and ovarian cancers, and hepatocellular carcinomas (6-9). A common germline mutation G84E in the HOXB13 protein has recently been found to be associated with significant increased risk of prostate cancer (10). Currently, HOXB13 is being evaluated as a marker for metastatic lesions of prostate origin (11,12).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: RalA and RalB are members of the Ras family of small GTPases and are highly homologous in protein sequence. The functions of RalA and RalB are distinct yet overlapping. By binding to various effector proteins, RalA and RalB serve as important GTP sensors for exocytosis and membrane trafficking (1-3). RalA is required for Ras-related tumorigenesis (4) and RalB is important for tumor survival (5). In addition to tumor formation, Ral proteins also play a role in cancer cell migration and metastatic tumor invasion (6,7).

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

Application Methods: Immunoprecipitation, Western Blotting

Background: DOT1-like protein (DOT1L), also known as Lysine N-methyltransferase 4 (KMT4), is a histone methyltransferase that functions to mono-, di-, and tri-methylate histone H3 on lysine 79, a histone modification that is associated with active transcription and plays a role in DNA damage response, cell cycle regulation, and embryonic stem cell development (1). DOT1L is required for the initiation and maintenance of mixed lineage leukemia (MLL)-rearranged leukemias, and selective DOT1L inhibitors such as EP2-5676 show remarkable anti-tumor effects in MLL-rearranged leukemias (2,3). Multiple studies have also implicated DOT1L in solid tumor cancers such as breast cancer, where DOT1L induces neoplastic transformation of immortalized breast cancer cells and promotes tumor initiation and growth. Overexpression of DOT1L is associated with poor prognosis in breast cancer, and selective DOT1L inhibitors are able to suppress proliferation and migration of breast cancer cells (4-6).

$348
100 µl
This Cell Signaling Technology antibody is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The HRP conjugated antibody is expected to exhibit the same species cross-reactivity as the unconjugated PKM2 (D78A4) XP® Rabbit mAb #4053.
APPLICATIONS
REACTIVITY
Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: Pyruvate kinase is a glycolytic enzyme that catalyses the conversion of phosphoenolpyruvate to pyruvate. In mammals, the M1 isoform (PKM1) is expressed in most adult tissues (1). The M2 isoform (PKM2) is an alternatively spliced variant of M1 that is expressed during embryonic development (1). Research studies found that cancer cells exclusively express PKM2 (1-3). PKM2 is shown to be essential for aerobic glycolysis in tumors, known as the Warburg effect (1). When cancer cells switch from the M2 isoform to the M1 isoform, aerobic glycolysis is reduced and oxidative phosphorylation is increased (1). These cells also show decreased tumorigenicity in mouse xenografts (1). Recent studies showed that PKM2 is not essential for all tumor cells (4). In the tumor model studied, PKM2 was found to be active in the non-proliferative tumor cell population and inactive in the proliferative tumor cell population (4).

$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 cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Stat2 (Tyr690) (D3P2P) Rabbit mAb #88410.
APPLICATIONS
REACTIVITY
Human, Rat

Application Methods: Flow Cytometry

Background: Stat2 (113-kDa), originally purified from the nuclei of alpha-interferon-treated cells, is critical to the transcriptional responses induced by type I interferons, IFN-alpha/beta (1,2). Knockout mice with a targeted disruption of Stat2 have higher susceptibility to viral infection and altered responses to type I interferons (3). Stat2 is rapidly activated by phosphorylation at Tyr690 in response to stimulation by IFN-alpha/beta via associations with receptor-bound Jak kinases (4). Unlike other Stat proteins, Stat2 does not form homodimers. Instead, activated Stat2 forms a heterodimer with Stat1 and translocates to the nucleus. There, it associates with the DNA-binding protein p48 and forms the transcriptional activator complex, interferon-stimulated gene factor 3 (ISGF3), promoting transcription from the ISRE (5).

$108
250 PCR reactions
500 µl
SimpleChIP® Human MDM2 Intron 2 Primers contain a mix of forward and reverse PCR primers that are specific to Intron 2 of the human double minute 2 protein (MDM2) 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®.
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: Western Blotting

Background: The Eph receptors are the largest known family of receptor tyrosine kinases (RTKs). They can be divided into two groups based on sequence similarity and on their preference for a subset of ligands. While EphA receptors bind to a glycosylphosphatidylinositol-anchored ephrin A ligand, EphB receptors bind to ephrin B proteins that have a transmembrane and cytoplasmic domain (1,2). Research studies have shown that Eph receptors and ligands may be involved in many diseases including cancer (3). Both ephrin A and B ligands have dual functions. As RTK ligands, ephrins stimulate the kinase activity of Eph receptors and activate signaling pathways in receptor-expressing cells. The ephrin extracellular domain is sufficient for this function as long as it is clustered (4). The second function of ephrins has been described as "reverse signaling", whereby the cytoplasmic domain becomes tyrosine phosphorylated, allowing interactions with other proteins that may activate signaling pathways in the ligand-expressing cells (5).The EphA1 receptor preferentially binds ephrin-A1 as a ligand (6). This ligand-receptor interaction stimulates EphA1 signaling and regulates cell morphology and motility through the ILK-RhoA-ROCK pathway (7). The EphA1 gene has been associated with late-onset Alzheimer's diseases (8,9). The role of EphA1 in cancer development falls into two opposite categories. In some type of cancer such as prostate, gastric and liver cancer, high expression of EphA1 associates with cancer metastasis and invasion (10-12). For other types of cancers, such as colon cancer and nonmelanoma skin cancer, down-regulation of the protein correlates with cancer development (13,14). The bidirectional signaling modulation of Ephrin-Ephrin receptor interaction might contribute this paradox phenomena (15).

$108
250 PCR reactions
500 µl
SimpleChIP® Human TRIO Exon 57 Primers contain a mix of forward and reverse PCR primers that are specific to exon 57 of the human trio rho guanine nucleotide exchange factor. 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®.
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

Application Methods: Immunoprecipitation, Western Blotting

Background: TROP2 is a transmembrane glycoprotein encoded by gene TACSTD2 (tumor associated calcium signal transducer 2). TROP2 was first discovered as a biomarker of invasive trophoblast cells and later reported in many types of cancer cells as well as in various organs during development, and adult stem cells during homeostasis (1, 2). TROP2 has an extracellular domain with EGF thyroglobulin type-1 repeats, a transmembrane domain and a short cytoplasmic tail with a HIKE domain containing a PIP2 binding site and PKC phosphoryaltion site (Ser303) (1-4). TROP2 functions by regulating multiple signaling pathways including-interaction of extracellular domain with integrin beta1 to regulate FAK signaling, association of its transmembrane domain with claudin1 and claudin7 for tight junction formation, as well as regulation of intracellular calcium release by its PIP2 binding and activation of the ERK/MAPK pathway (1,2, 5-8). All of these functions are important for its role in tumor proliferation, metastasis and invasion (1,2). PKC can phosphorylate TROP2 at Ser303; the phosphorylation changes the cytoplasmic tail conformation and further promotes its signaling (9). TROP2 can be activated through intramembrane proteolysis first by TACE, followed further cleavage by presenilin 1 and presenilin2. The proteolysis process is required for its role in tumor cell proliferation (10,11).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Monkey

Application Methods: Immunohistochemistry (Paraffin), Western Blotting

Background: Neural cell adhesion molecule L1 (NCAM-L1/L1CAM) is a single pass transmembrane glycoprotein member of the immunoglobulin superfamily, containing six amino-terminal extracellular Ig-like domains followed by five fibronectin type-III domains (1). NCAM-L1 is mainly expressed in the brain, and plays an important role in the developing nervous system, with involvement in neurite fasciculation and outgrowth, myelination, neuronal migration, and neuronal cell adhesion (2). Mutations in the NCAM-L1 gene cause varying degrees of neurological disease including X-linked hydrocephalus, MASA syndrome, spastic paraplegia type 1, and X-linked corpus callosum agenesis, together known as L1 syndrome (3). Apart from the nervous system, NCAM-L1 is overexpressed in many cancers and supports a poor prognosis by facilitating aggressive tumor growth, metastasis and chemoresistance (4,5).

$364
50 tests
100 µl
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye 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 Phospho-SLP-76 (Ser376) (D7S1K) XP® Rabbit mAb #92711.
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry

Background: SH2 domain-containing leukocyte protein of 76 kDa (SLP-76) is a hematopoietic adaptor protein that is important in multiple biochemical signaling pathways and necessary for T cell development and activation (1). ZAP-70 phosphorylates SLP-76 and LAT as a result of TCR ligation. SLP-76 has amino-terminal tyrosine residues followed by a proline rich domain and a carboxy-terminal SH2 domain. Phosphorylation of Tyr113 and Tyr128 result in recruitment of the GEF Vav and the adapter protein Nck (2). TCR ligation also leads to phosphorylation of Tyr145, which mediates an association between SLP-76 and Itk, which is accomplished in part via the proline rich domain of SLP-76 and the SH3 domain of ITK (3). Furthermore, the proline rich domain of SLP-76 binds to the SH3 domains of Grb2-like adapter Gads (3,4). In resting cells, SLP-76 is predominantly in the cytosol. Upon TCR ligation, SLP-76 translocates to the plasma membrane and promotes the assembly of a multi-protein signaling complex that includes Vav, Nck, Itk and PLCγ1 (1). The expression of SLP-76 is tightly regulated; the protein is detected at very early stages of thymocyte development, increases as thymocyte maturation progresses, and is reduced as cells mature to CD4+ CD8+ double-positive thymocytes (5).

$84
100 µl
Ghost Dye™ Red 710 Viability Dye is used to discriminate viable from non-viable mammalian cells in flow cytometry applications. Ghost Dye™ Red 710 Viability Dye irreversibly binds free amines available on the cell surface as well as intracellular free amines exposed in cells with compromised cell membranes. Non-viable cells with loss of membrane integrity will react with significantly more Ghost Dye™ Red 710 Viability Dye than healthy cells in the same sample. Cells that exhibit increased fluorescence intensity can be excluded from analysis.
APPLICATIONS

Application Methods: Flow Cytometry

$303
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 by ATM/ATR, followed by autophosphorylation of Ser296. Activation occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for re-entry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of aurora B and BubR1 (8). Research studies have implicated Chk1 as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).