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Product listing: Phospho-Tyrosine Hydroxylase (Ser31) Antibody, UniProt ID P04177 #3370 to SMYD2 Antibody, UniProt ID Q9NRG4 #4251

$303
100 µl
APPLICATIONS
REACTIVITY
Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Tyrosine hydroxylase (TH) catalyzes the rate-limiting step in the synthesis of the neurotransmitter dopamine and other catecholamines. TH functions as a tetramer, with each subunit composed of a regulatory and catalytic domain, and exists in several different isoforms (1,2). This enzyme is required for embryonic development since TH knockout mice die before or at birth (3). Levels of transcription, translation and posttranslational modification regulate TH activity. The amino-terminal regulatory domain contains three serine residues: Ser9, Ser31 and Ser40. Phosphorylation at Ser40 by PKA positively regulates the catalytic activity of TH (4-6). Phosphorylation at Ser31 by CDK5 also increases the catalytic activity of TH through stabilization of TH protein levels (7-9).

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

Application Methods: Western Blotting

Background: LIS1 is a cytoskeleton-interacting protein that contains an N-terminal dimerization domain and a C-terminal β-propeller domain that interacts with the motor domain of dynein (1-3). Research studies have shown that mutations in the LIS1 gene are involved in lissencephaly, a disease characterized by severe defects in brain development (4). LIS1 also plays a critical role in cortical migration and development in the brain (5). LIS1 activity is required for retrograde translocation of excitatory synapses in developing interneuron dendrites in a microtubule-dependent fashion (6).

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

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: RCC2/TD-60 is a member of the RCC1 (regulator of chromosome condensation 1) family of guanine nucleotide exchange factors. RCC2/TD-60 is associated with the chromosome passenger complex (CPC), which also consists of aurora B kinase, borealin, INCENP (inner centromere protein) and survivin. The CPC acts at various stages of mitosis, interacts with microtubules and is required for proper chromosome segregation and cytokinesis. Regulation of aurora B kinase is key in the regulation of the CPC (reviewed in 1,2). In late mitosis, RCC2/TD-60 is required for spindle assembly and recruitment of survivin and aurora B (3). RCC2/TD-60 is also required for aurora B activation in vitro and localization of the CPC to centromeres (4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Flow Cytometry, Immunofluorescence (Immunocytochemistry), Immunoprecipitation

Background: Spi-B belongs to the ETS family of transcription factors and activates transcription through direct binding to the PU box (1). Spi-B was identified based on its homology with PU.1/Spi-I (1). The function of Spi-B is most well studied in B cells and plasmacytoid dendritic cells (pDCs). B cells from mice deficient in Spi-B proliferate poorly and die in response to signaling through the B cell receptor (2). In addition, Spi-B(-/-) mice have a defect in germinal center formation (2). Spi-B plays a role in both the development and function of pDCs. It is highly expressed by pDCs and not by monocyte-derived DCs, and overexpression of Spi-B in hematopoietic progenitor cells inhibits development of other lymphoid lineages (3). pDCs from Spi-B(-/-) mice produce less interferon in response to TLR7 and TLR9 stimulation and have lower expression of pDC-specific surface markers (4). Finally, Spi-B is required for differentiation of intestinal microfold cells (5,6).

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

Application Methods: Immunoprecipitation, Western Blotting

Background: The flightless-I (fliI) gene was first identified in Drosophila mutant screens for genes involved in flight behavior. Homozygous mutant alleles at the fliI locus are embryonic lethal, whereas heterozygous mutations yield a "flightless" phenotype resulting from defects in flight muscle fiber development (1). The encoded protein (flightless-I, FLII) is a highly conserved member of the gelsolin superfamily, defined by the presence of C-terminal gelsolin motifs that function as actin-binding domains (2). Genetic knock-out studies in mice and worms confirmed that Flightless-I plays a critical and highly conserved role in embryonic development, likely through its effects on actin remodeling of the cytoskeleton (3,4). Postnatally, Flightless-I is recognized to play an important role in wound repair (5). Flightless-I protein levels are increased in many wound types, and depletion of Flightless-I protein levels has been shown to accelerate wound repair by promoting fibroblast proliferation and epithelial migration (6-8). Studies in animal models suggest that Flightless-I may inhibit the wound repair process by modulating TGF-β signaling dynamics in the wound environment (9).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: Heterotrimeric guanine nucleotide-binding proteins, G proteins, transduce ligand binding to G protein-coupled receptors (GPCRs) into intracellular responses (1). G proteins are comprised of 3 subunits, alpha (Gα), beta (Gβ), and gamma (Gγ). Upon activation of GPCRs, the receptor promotes the exchange of GDP to GTP of Gα, changing the confirmation of the switch regions within Gα. The receptor bound heterotimeric G protein (inactive) is then released, and dissociates into the GTP-bound Gα (active) monomer and the Gβ/Gγ heterodimer (1,2). Gα activates adenylyl cyclase, which converts ATP to the second messenger cAMP. Gα also activates phosphoinositide-specific phospholipase C (PLC), which catalyzes hydrolysis of the phospholipid of phosphatidylinositol 4,5-biphosphate (PIP2), releasing the second messengers IP3 and 1,2-diacylglycerol (DAG). IP3 activates IP3 receptors to release Ca2+ from the ER. DAG is an activator of protein kinase C (PKC), which in turn activates the Erk1/2 pathway (1,3). The primary function of the Gβ/Gγ heterodimer is to inhibit Gα, although it may also activate second messengers (e.g. PLC pathway) or gate ion channels (e.g. GIRK) (1). Guanine nucleotide-binding protein b3 (GNB3) is an isoform of the b subunit. Research studies have shown that a polymorphism in the GNB3 gene, C825T, is associated with hypertension, obesity, and depression (4).

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

Application Methods: Immunoprecipitation, Western Blotting

Background: Heterogeneous nuclear ribonucleoprotein Q and R belong to a family of hnRNP proteins that are involved in RNA binding, RNA biosynthesis, and mRNA transport from the nucleus to the cytoplasm (1-3). These two proteins are encoded by different genes but have 83% homology. hnRNP Q has three alternative splice variants (hnRNP Q1-3) (1-3). Methylation of carboxy-terminal arginine residues is required for nuclear localization (4). hnRNP Q binds to AU-rich mRNA in conjunction with AUF1 and regulates mRNA decay (5). hnRNP Q isoforms play a crucial role in mediating nuclear function of survival of motor neuron (SMN) complex (6,7) and modulating RNA biosynthesis and hepatitis C virus replication (8). hnRNP R was identified recently and its function is still under investigation (9), however hnRNP R does not duplicate the biological function of hnRNP Q. Both hnRNP Q and R are present in cytoplasmic mRNP granules containing untranslated mRNAs (10) and both interact with SMN (11).

$262
3 nmol
300 µl
SignalSilence® p70/85 S6 Kinase siRNA I allows the researcher to specifically inhibit p70/85 S6 kinase expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.
REACTIVITY
Human

Background: p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8).

$115
100 µl
The extract is prepared from whole heart tissue of adult mice, and is intended for use as a positive control in western blotting applications. The protein concentration is 2 mg/ml.The extract was prepared from whole tissue by homogenization in 1X RIPA buffer (#9806, 10X) (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 1 mM sodium EDTA, 1 mM EGTA, 1 μg/ml leupeptin, 1 mM β-glycerophosphate, 1 mM sodium orthovanadate, 2.5 mM sodium pyrophosphate), 1 mM PMSF (#8553, 34.84 mg), 1X Protease/Phosphatase Inhibitor Cocktail (#5872, 100X).The extract was subsequently sonicated and insoluble cell debris removed by centrifugation. The extract was then boiled for 5 min in 1X SDS sample buffer + DTT (#7722, Blue Loading Buffer Pack) (62.5 mM Tris-HCl pH 6.8, 10% glycerol, 2% SDS, 0.01% bromophenol blue, 42 mM DTT) to denature the proteins.
APPLICATIONS

Application Methods: Western Blotting

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Peroxiredoxin 6 (PRDX6) belongs to an antioxidant enzyme family of non-seleno peroxidases. PRDX6 is a unique member of the PRDX family, exhibiting both glutathione peroxidase and phospholipase A2 activities (1,2). PRDX6 regulates phospholipid turnover in addition to protecting cells against oxidative injury. PRDX6 is expressed in all major organs, with a particularly high level in lung, where it regulates lung surfactant phospholipid synthesis and turnover (3-5). Research studies have shown that PRDX6 is aberrantly expressed in various cancers, and can promote cancer cell metastasis and invasion (6,7). Elevated expression of PRDX6 and related PRDX family members has also been shown to contribute to drug resistance in cancer cells (8,9). PRDX6 is also expressed in neutrophils, where it has been shown to activate NADPH oxidase (NOX2) (10-12).

SignalSilence® Akt siRNA II from Cell Signaling Technology allows the researcher to specifically inhibit Akt expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products are rigorously tested in-house and have been shown to reduce protein expression by western analysis.

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
Mouse

Application Methods: Immunofluorescence (Frozen)

Background: Vasopressin is a neuroendocrine peptide that is released to the circulation by magnocellular neurons whose cell bodies are mainly found in the paraventricular and the supraoptic nuclei of the hypothalamus. It was first isolated from pituitary gland extracts and synthesized in 1951 (1). Vasopressin acts by activating G protein-coupled, V1a, V1b (also known as V3) and V2 receptors and plays a fundamental role in the maintenance of water homeostasis. One of its main functions is body water retention (2), hence its alternative name antidiuretic hormone or ADH. Vasopressin also leads to increased arterial blood pressure by raising peripheral vascular resistance (3). Vasopressin is also involved in other physiological processes such as acute heart failure (4), pain (5), and metabolic syndrome (6).

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

Application Methods: Western Blotting

Background: Initiation of eukaryotic DNA replication is a stringently regulated process that requires the cooperation of many proteins and protein complexes to occur efficiently, at the origins of replication, and once per cell cycle. The initiation of DNA replication requires a protein complex composed of two DNA polymerase α subunits and a pair of primase subunits. Primase activity catalyzes de novo synthesis of an RNA/DNA primer (initiator DNA) on the leading and lagging strands, while polymerase activity extends the initiator DNA (1). The 48 and 58 kDa primase subunits cooperate in the synthesis of small RNA primers. p48 is the catalytically active subunit (2), while p58 couples p48 to the polymerase to allow the transfer of primers to the active site. The p58 subunit may also play a role in regulation of primer length (3,4).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: The suppressor of cytokine signaling (SOCS) family members are negative regulators of cytokine signal transduction that inhibit the Jak/Stat pathway (1-3). The SOCS family consists of at least 8 members including the originally identified cytokine-inducible SH2-containing protein (CIS1), as well as SOCS1-7. Each SOCS family member contains a central SH2 domain and a conserved carboxy-terminal motif designated as the SOCS box. These proteins are important regulators of cytokine signaling, proliferation, differentiation, and immune responses.

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: The structural maintenance of chromosomes 2 (SMC2) and 4 (SMC4) proteins are condensin complex subunits that enable chromosome condensation and compaction during migration to opposite poles during anaphase (1,2). Condensin is a general regulator of chromosome architecture that may also regulate gene expression and DNA repair. Condensin complex subunits SMC2 and SMC4 form a functional ATPase essential for chromatin condensation, while three auxiliary subunits regulate ATPase activity. Both SMC2 and SMC4 are found within two distinct condensin complexes (condensin I and II) in higher eukaryotes. Condensin I contains auxiliary subunits NCAPD2, NCAPG, and NCAPH, while condensin II contains related auxiliary proteins NCAPD3, NCAPG2, and NCAPH2 (1,2).Each condensin complex exhibits different localization patterns during the cell cycle and provides for distinct functions during mitosis (3-5). Condensin I is cytoplasmic during interphase and binds chromatin following the breakdown of the nuclear envelope at the end of prophase. Condensin I is required for complete dissociation of cohesin from chromosome arms, for chromosome shortening, and for normal timing of progression through pro-metaphase and metaphase. Mutations in corresponding condensin I genes result in cytokinesis defects due to the persistence of anaphase fibers. Condensin II is nuclear during interphase, but does not bind to chromatin until early prophase where it remains bound until the end of telophase. Condensin II is required for initial chromatin condensation during early prophase. Mutations in corresponding condensin II genes produce high numbers of anaphase bridges resulting from incomplete chromosome segregation. Condensin II complex subunit D3 (NCAPD3) plays a pivotal role in the loading of condensin II onto chromatin and the regulation of chromatin condensation (6,7). NCAPD3 protein contains HEAT repeat clusters that bind to mono-methyl histone H4 Lys20, a histone mark prevalent during mitosis and important for DNA repair and chromatin condensation (6). Increased mono-methyl histone H4 Lys20 levels caused by dissociation of the histone demethylase PHF8 from chromatin and increased expression of the methyltransferase SET8, leads to increased binding of NCAPD3 and condensin II to chromosomes early in mitosis (6). Phosphorylation of NCAPD3 at Thr1415 by CDK1 kinase (cdc2) leads to the recruitment of PLK1 kinase, which hyperphosphorylates condensin II and facilitates mitotic chromosome assembly (7).

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

Application Methods: Western Blotting

Background: The 26S proteasome is a highly abundant proteolytic complex involved in the degradation of ubiquitinated substrate proteins. It consists largely of two sub-complexes, the 20S catalytic core particle (CP) and the 19S/PA700 regulatory particle (RP) that can cap either end of the CP. The CP consists of two stacked heteroheptameric β-rings (β1-7) that contain three catalytic β-subunits and are flanked on either side by two heteroheptameric α-rings (α1-7). The RP includes a base and a lid, each having multiple subunits. The base, in part, is composed of a heterohexameric ring of ATPase subunits belonging to the AAA (ATPases Associated with diverse cellular Activities) family. The ATPase subunits function to unfold the substrate and open the gate formed by the α-subunits, thus exposing the unfolded substrate to the catalytic β-subunits. The lid consists of ubiquitin receptors and DUBs that function in recruitment of ubiquitinated substrates and modification of ubiquitin chain topology (1,2). Other modulators of proteasome activity, such as PA28/11S REG, can also bind to the end of the 20S CP and activate it (1,2).

$305
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 Itk (2F12) Mouse mAb #2380.
APPLICATIONS
REACTIVITY
Human

Application Methods: Western Blotting

Background: Interleukin-2 inducible T-cell kinase (Itk, Emt or Tsk) is a member of the non-receptor protein tyrosine kinases. Family members of Itk include Tec, Btk, Rlk and Bmx and are all defined by a common structure: an amino-terminal PH domain, a Tec-homology domain and a SH3 and SH2 domain followed by a carboxy-terminal kinase domain (1). Tec, Rlk and Itk are expressed in T cells and activated in response to T cell receptor (TCR) engagement. Data demonstrate that Itk functions in signal transduction downstream of TCR and activates PLCgamma1 and Erk. Lck directly activates Itk through phosphorylation in the conserved activation loop at Tyr511, and furthermore, Itk is autophosphorylated in the SH3 domain at Tyr180. Itk-Y180F is still capable of phosphorylating PLCgamma1 in contrast to Itk-Y511F, which has lost that function (2-3). Itk -/- mice show reduced lung inflammation, eosinophil infiltration and mucous production in response to allergic asthma induction. Thus, Itk could become a desirable target for anti-asthmatic treatments (4).

$303
100 µl
APPLICATIONS
REACTIVITY
Rat

Application Methods: Western Blotting

Background: N-methyl-D-aspartate receptor (NMDAR) forms a heterodimer of at least one NR1 and one NR2A-D subunit. Multiple receptor isoforms with distinct brain distributions and functional properties arise by selective splicing of the NR1 transcripts and differential expression of the NR2 subunits. NR1 subunits bind the co-agonist glycine and NR2 subunits bind the neurotransmitter glutamate. Activation of the NMDA receptor or opening of the ion channel allows flow of Na+ and Ca2+ ions into the cell, and K+ out of the cell (1). Each subunit has a cytoplasmic domain that can be directly modified by the protein kinase/phosphatase (2). PKC can phosphorylate the NR1 subunit (NMDAR1) of the receptor at Ser890/Ser896, and PKA can phosphorylate NR1 at Ser897 (3). The phosphorylation of NR1 by PKC decreases its affinity for calmodulin, thus preventing the inhibitory effect of calmodulin on NMDAR (4). The phosphorylation of NR1 by PKA probably counteracts the inhibitory effect of calcineurin on the receptor (5). NMDAR mediates long-term potentiation and slow postsynaptic excitation, which play central roles in learning, neurodevelopment, and neuroplasticity (6).

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

Application Methods: Chromatin IP, Immunoprecipitation, Western Blotting

Background: The E2F family consists of 8 transcription factors that regulate genes that control cell cycle progression by complexing with DP and Rb proteins (1-4). E2F transcriptional activation is generally opposed by associating with RB proteins, pRB, p107, and p130 (5-7). E2F-1, -2, and -3a function as activators that can help quiescent cells enter S phase, while E2F-3b, -4, and -5 repress cell growth through the recruitment of HDAC’s and other corepressors to target genes (8-10). E2F-6 diverges considerably from other family members, and has repressive properties governed not through interaction with Rb proteins, but by recruiting the polycomb repressive complex (11,12). E2F-7, and -8 are unique in that they have two DNA-binding domains and do not heterodimerize with DP proteins. These E2F family members repress transcription and delay progression of the cell cycle through the regulation of E2F-1 (13-15)

The BAF Complex Antibody Sampler Kit II provides an economical means of detecting levels of various BAF complex components. The kit contains enough primary antibodies to perform at least two western blot experiments.
$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Western Blotting

Background: CASK is an adaptor protein with a calcium/calmodulin-dependent protein kinase domain, an SH3 domain, a guanylate kinase homology domain (GUK), and a PDZ domain. CASK links transmembrane proteins to the cytoskeleton and signaling molecules. In particular, CASK binds to neurexin to stabilize pre- and post-synaptic structures (1). While most CASK protein (~80%) is cytoplasmic, a portion of the protein enters the nucleus, where it acts as a transcriptional coactivator (2). Transgenic mice with CASK insertional mutations die within 24 hours of birth (3).

$260
100 µl
APPLICATIONS
REACTIVITY
Human, Mouse, Rat

Application Methods: Immunoprecipitation, Western Blotting

Background: Synaptophysin (SYP) is a neuronal synaptic vesicle glycoprotein that is expressed in neuroendocrine cells and neoplasms (1). Synaptophysin contains four transmembrane domains that form a hexameric channel or gap junction-like pore (2). Synaptophysin binds to the SNARE protein synaptobrevin/VAMP, which prevents the inclusion of synaptobrevin in the synaptic vesicle fusion complex and creates a pool of synaptobrevin for exocytosis when synapse activity increases (3). Synaptophysin is also responsible for targeting synaptobrevin 2/VAMP2 to synaptic vesicles, a critical component of the fusion complex (4).

$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 antibody YAP (D8H1X) XP® Rabbit mAb #14074.
APPLICATIONS
REACTIVITY
Hamster, Human, Monkey, Mouse, Rat

Application Methods: Western Blotting

Background: YAP (Yes-associated protein, YAP65) was identified based on its ability to associate with the SH3 domain of Yes. It also binds to other SH3 domain-containing proteins such as Nck, Crk, Src, and Abl (1). In addition to the SH3 binding motif, YAP contains a PDZ interaction motif, a coiled-coil domain, and WW domains (2-4). While initial studies of YAP all pointed towards a role in anchoring and targeting to specific subcellular compartments, subsequent studies showed that YAP is a transcriptional co-activator by virtue of its WW domain interacting with the PY motif (PPxY) of the transcription factor PEBP2 and other transcription factors (5). In its capacity as a transcriptional co-activator, YAP is now widely recognized as a central mediator of the Hippo Pathway, which plays a fundamental and widely conserved role in regulating tissue growth and organ size. Phosphorylation at multiple sites (e.g., Ser109, Ser127) by LATS kinases promotes YAP translocation from the nucleus to the cytoplasm, where it is sequestered through association with 14-3-3 proteins (6-8). These LATS-driven phosphorylation events serve to prime YAP for subsequent phosphorylation by CK1δ/ε in an adjacent phosphodegron, triggering proteosomal degradation of YAP (9).

$489
96 assays
1 Kit
The PathScan® Total SQSTM1/p62 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total SQSTM1/p62 protein. An SQSTM1/p62 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, SQSTM1/p62 protein is captured by the coated antibody. Following extensive washing, SQSTM1/p62 Mouse Detection Antibody is added to detect the captured SQSTM1/p62 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 absorbance for this developed color is proportional to the quantity of total SQSTM1/p62 protein.Antibodies in kit are custom formulations specific to kit.
REACTIVITY
Human, Mink, Monkey, Mouse, Rat

Background: Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein involved in cell signaling, oxidative stress, and autophagy (1-4). It was first identified as a protein that binds to the SH2 domain of p56Lck (5) and independently found to interact with PKCζ (6,7). SQSTM1 was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins and triggering degradation of proteins through the proteasome or lysosome (8). Interaction between SQSTM1 and TRAF6 leads to the K63-linked polyubiquitination of TRAF6 and subsequent activation of the NF-κB pathway (9). Protein aggregates formed by SQSTM1 can be degraded by the autophagosome (4,10,11). SQSTM1 binds autophagosomal membrane protein LC3/Atg8, bringing SQSTM1-containing protein aggregates to the autophagosome (12). Lysosomal degradation of autophagosomes leads to a decrease in SQSTM1 levels during autophagy; conversely, autophagy inhibitors stabilize SQSTM1 levels. Studies have demonstrated a link between SQSTM1 and oxidative stress. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (3). Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity.

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

Application Methods: Western Blotting

Background: The 26S proteasome is a highly abundant proteolytic complex involved in the degradation of ubiquitinated substrate proteins. It consists largely of two sub-complexes, the 20S catalytic core particle (CP) and the 19S/PA700 regulatory particle (RP) that can cap either end of the CP. The CP consists of two stacked heteroheptameric β-rings (β1-7) that contain three catalytic β-subunits and are flanked on either side by two heteroheptameric α-rings (α1-7). The RP includes a base and a lid, each having multiple subunits. The base, in part, is composed of a heterohexameric ring of ATPase subunits belonging to the AAA (ATPases Associated with diverse cellular Activities) family. The ATPase subunits function to unfold the substrate and open the gate formed by the α-subunits, thus exposing the unfolded substrate to the catalytic β-subunits. The lid consists of ubiquitin receptors and DUBs that function in recruitment of ubiquitinated substrates and modification of ubiquitin chain topology (1,2). Other modulators of proteasome activity, such as PA28/11S REG, can also bind to the end of the 20S CP and activate it (1,2).

$131
200 µl
This Cell Signaling Technology antibody is conjugated to FITC under optimal conditions and tested in-house for direct flow cytometric analysis in human cells.
APPLICATIONS

Application Methods: Flow Cytometry

Background: Isotype control antibodies are used to estimate the nonspecific binding of target primary antibodies due to Fc receptor binding or other protein-protein interactions. An isotype control antibody should have the same immunoglobulin type and be used at the same concentration as the test antibody.

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunoprecipitation, Western Blotting

Background: In mammalian cells, the significance of histone H2B ubiquitination in chromatin epigenetics came from the identification of the budding yeast protein Bre1 (1,2). Together with the ubiquitin-conjugating enzyme Rad6, Bre1 serves as the E3 ligase in the monoubiquitination of the yeast histone H2B within transcribed regions of chromatin (1-3). Subsequently, the mammalian orthologs of yeast Bre1, RNF20 and RNF40, were identified (4,5). These two proteins form a tight heterodimer that acts as the major E3 ligase responsible for histone H2B monoubiquitination at Lys120 in mammalian cells, a modification linked to RNA Pol II-dependent transcription elongation in undamaged cells. Researchers have shown that DNA double-strand breaks (DSBs) are also capable of inducing monoubiquitination of H2B. This process depends upon the recruitment to DSB sites, as well as ATM-dependent phosphorylation of the RNF20-RNF40 heterodimer, thus highlighting a role for this E3 ligase in DSB repair pathways (6). Indeed, investigators have shown that loss of RNF20-RNF40 function promotes replication stress and chromosomal instability, which may constitute an early step in malignant transformation that precedes cell invasion (7).

$260
100 µl
APPLICATIONS
REACTIVITY
Human

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: The RecQ family is a group of DNA helicases that play an important role in global genomic stability (1). Mutations in three of the five known human RecQ proteins (BLM, WRN and RECQL4) give rise to clinically distinct disorders that are characterized by features such as premature aging and predisposition to cancer (2,3). The clinical distinction of each disease associated with these mutations points to distinct roles that members of this helicase family play in DNA metabolism. RecQL1 is the most abundant protein of the RecQ family and was the first family member to be discovered. No disease associations have been reported with RecQL1 and its biological activities are not well understood (4). It has recently been shown that depletion of RecQL1 negatively affects genomic maintenance and cellular proliferation – which may point to a role in DNA damage repair and cell cycle progression (5,6). Upregulation of RecQL1 along with other RecQ family members has been reported in cells in response to oncogenic viral infection (7).

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

Application Methods: Western Blotting

Background: SET and MYND domain-containing protein 2 (SMYD2), also known as lysine methyltransferase protein 3C (KMT3C), is a member of the SMYD family of protein methyltransferases (1). All five members of this family (SMYD1, SMYD2, SMYD3, SMYD4, and SMYD5) contain a conserved catalytic SET domain, originally identified in Drosophila Su[var]3-9, Enhancer of zeste, and Trithorax proteins. This domain is split by the MYN domain/zinc finger motif believed to facilitate protein-protein interactions (1). SMYD2 localizes to both the cytoplasm and nucleus, and is highly expressed in the adult mouse heart, brain, liver, kidney, thymus, and ovary, as well as in the developing mouse embryo (1). SMYD2 functions to repress transcription by interacting with the Sin3A repressor complex and methylating Lys36 of histone H3 (1). SMYD2 also interacts with HSP90α and methylates Lys4 of histone H3, a mark associated with transcriptional activation (2). In addition to histones as methyl substrates, SMYD2 methylates p53 at Lys370 to repress p53-mediated transcriptional activation and apoptosis (3,4).