- 中文名称
Glycogen Synthase Kinase 3 beta, Total, Human (GSK3b) BioAssay ELISA Kit
- 英文名字
- Glycogen Synthase Kinase 3 beta, Total, Human (GSK3b) BioAssay ELISA Kit
- 供应商
- USBiological
- 产品货号
- G8170-60
- 产品报价
- ￥1/96Tests
- 产品说明书
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- 产品新闻
- 背景资料
- USBiological品牌是全球有名的抗原抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。武汉艾美捷科技作为USBiological品牌中国区域总代理，是行业中少有的致力于服务客户，帮助客户，且拥有独立的专业销售团队、技术支持团队、市场营销团队、进出口报关团队的高科技生物企业。可以为您提供及时的咨询响应，专业的产品和解决方案支持，稳健快捷的交货周期，优质放心的售后服务。我们致力于为您提供有价值的产品和服务，在意您的成功！
- 产品描述
- Glycogen synthase kinase-3b (GSK-3b), a serine/threonine protein kinase, is one of two mammalian GSK-3 isoforms, designated GSK-3a (Mr=51kD) and GSK-3b (Mr=47kD). The two isoforms map to different genes: in humans, GSK-3a maps to 19q13.2, while GSK-3b maps to 3q13.3. The kinase domains of the two isoforms share the greatest homology (approximately 95% homology), while the C-terminal regions are found to diverge between the two isoforms (36% homology). The two isoforms also differ slightly in structure. GSK-3a contains a glycine-rich extension in its N-terminus that is absent from GSK-3b, a feature that accounts for the difference in the molecular masses of the two isoforms. GSK-3b homologs appear to be ubiquitously expressed in eukaryotes. The Drosophila homolog is designated Shaggy/Zeste-white3. The GSKs were initially identified as negative regulators of glycogen synthase, a key enzyme in glycogen metabolism. Recent studies have revealed that GSK-3b also phosphorylates numerous other substrates, including b-catenin, CREB, p53, c-Myc, c-Jun, heat shock factor-1, cyclin D1, Bax, axin, the adenomatous polyposis coli (APC) gene product, eukaryotic protein synthesis initiation factor 2B (eIF-2B) and the microtubule associated proteins MAP1B and tau, suggesting important roles for this kinase in diverse cell functions.GSK-3b is phosphorylated at several sites. Phosphorylation of tyrosine 216, located in GSK-3b’s activation loop, activates the enzyme, but may not be strictly required. Stimuli that enhance phosphorylation at tyrosine 216 include: staurosporine treatment, neurotrophic factor withdrawal with neuronal cell lines, lysophosphatidic acid (LPA) acting through the G proteins Ga12 and Ga13 and Ca2+ flux. Recent evidence implicate both Fyn and PYK2 in the phosphorylation of tyrosine 216. Phosphorylation of serine 9, located in the GSK-3b’s N-terminal pseudosubstrate domain, inhibits enzyme activity. Phosphorylation of serine 9 produces a primed pseudosubstrate that binds to a positively charged pocket formed by arginine 96, arginine 180 and lysine 205 and competes with protein substrates. Stimuli that enhance phosphorylation of serine 9 include insulin, EGF, PDGF (through MAPKAP-K1), amino acids (through p70S6K), PKA activators and PKC activators. Analysis of substrate specificity indicates that GSK-3b’s consensus sequence is Ser/Thr-X-X-X-Ser/Thr-P, where the N-terminal serine or threonine residue is the target for phosphorylation, X is any amino acid residue, but is often found to be proline and the C-terminal serine or threonine is a residue that has undergone a priming phosphorylation by a different enzyme activity, such as PKA or CKI. This priming phosphorylation enhances the efficiency of phosphorylation of the target residues by GSK-3b by as much as 1000 fold. Crystallographic studies have revealed that the phosphorylated residue of primed substrates binds to GSK-3b’s positively charged pocket, orienting the substrate within GSK-3b’s catalytic groove. Consistent with this model, negatively charged residues can substitute for the phosphorylated serine or threonine of the primed substrate in enhancing the catalytic efficiency of GSK-3b.GSK-3b has been implicated in several signaling pathways, including the insulin, Wnt and Hedgehog signaling pathways. In response to insulin, PI3K and AKT/PKB are activated and in turn inhibit GSK-3b by enhancing phosphorylation at serine 9. The result is decreased phosphorylation of GSK-3b substrates including glycogen synthase and eIF-2B and increased glycogen synthesis and protein synthesis. In Wnt signaling, GSK-3b is inhibited in a pathway that involves the protein, Dishevelled. Under resting conditions, active GSK-3b phosphorylates CKI-primed b-catenin, an event that targets b-catenin for ubiquitinylation and degradation by the proteasome. With the inactivation of GSK-3b in response to Wnt, b-catenin accumulates and translocates to the nucleus, where it influences gene expression. Hedgehog signaling similarly includes alterations in proteolysis of a GSK-3b consensus sequence-containing signaling protein designated Gli (the mammalian homolog of Cubitus interruptus [Ci] in Drosophila). As with b-catenin in the Wnt pathway, Gli also appears to play a role in transmitting signals into the cell nucleus. GSK-3b is currently under investigation in several important areas of research. Its interaction with the Wnt and Hedgehog pathways suggests a role in cell fate and morphology and possibly the etiology of certain cancers. Its role in transducing the insulin signal has made it a target for study in type II diabetes. GSK-3b is also under investigation in several neurological disorders. GSK-3b is highly expressed in neurons and is found to phosphorylate the microtubule binding protein tau at sites that are abnormally phosphorylated in Alzheimer’s disease. GSK-3b is also implicated in schizophrenia and bipolar disorder. GSK-3b (Total) ELISA is designed to detect and quantify the level of GSK-3b protein. This assay is intended for the detection of GSK-3b from lysates of human, mouse and rat cells. This kit can be used to normalize the phosphorylated GSK-3b content of the samples when using the GSK-3b [pS9] ELISA kit. This kit has been configured for research use only and is not to be used in diagnostic procedures.Principle of the Method:The GSK-3b (Total) kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibodyspecific for GSK-3b (regardless of phosphorylation state) has been coated onto the wells of the microtiter strips provided. Samples, including a standard containing GSK-3b (Total), control specimens, and unknowns, are pipetted into these wells. During the first incubation, the GSK-3b (Total) antigen binds simultaneously to the immobilized (capture) antibody and to the solution phase rabbit polyclonal (detection) antibody specific for GSK-3b. After washing, a horseradish peroxidase-labeled anti-rabbit IgG (anti-rabbit IgG-HRP) is added. This binds to the detection antibody to complete the four-member sandwich. After the second incubation and washing to remove the excess anti-rabbit IgG-HRP, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of GSK-3b (Total) present in the original specimen.Kit Components:G8170-60A: Microtiter Plate, 1x96wells. GSK-3b antibody-coated wells.*G8170-60B: Standard. 2x30ng. Reconstitute with 1.2ml of Standard Diluent Buffer G8170-60C.G8170-60C: Standard Diluent Buffer, 1x25ml. Contains 15 mM sodium azide, red dye* G8170-60D: GSK-3b Pab, 1x6ml. Contains 15 mM sodium azide, blue dye*G8170-60E: IgG (HRP), 100X, 1x125ul. Contains 3.3mM thymol.G8170-60F: HRP Diluent, 1x25ml. Contains 3.3mM thymol, yellow dye*G8170-60G: Wash Buffer Concentrate, 25X, 1x100ml.G8170-60H: TMB, 1x25ml.G8170-60J: Stop Solution, 1x25ml.Storage and Stability: Store at 4°C. Store G8170-60B powder at 4°C liquid at -70°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
- 产品特点
- 针对Glycogen Synthase Kinase 3 beta, Total, Human (GSK3b) BioAssay ELISA Kit该产品的特点优势，欢迎查阅官网提供的产品说明书。
- 保存建议
- 建议收到Glycogen Synthase Kinase 3 beta, Total, Human (GSK3b) BioAssay ELISA Kit产品后将其置于4°C/-70°C保存。
- 其他
- Usbiological公司是美国著名的抗体和生化试剂供应商，生产世界上种类最多的抗体，用于Western Blot、免疫沉淀、免疫荧光、免疫组化和流式细胞术等多种检测方法。Usbiological公司现已拥有超过50,19234种抗体、抗原和生化产品，为科研用户提供了诸多超值选择。
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- 注意
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