GSK3B (基因名), Glycogen synthase kinase-3 beta (蛋白名), GSK3B_HUMAN.
产品名称:
Human GSK3B/ Glycogen synthase kinase-3 beta ELISA Kit
糖原合成酶激酶3β
货号:
E2298h
商标:
EIAab®
监管等级:
别名:
Serine/threonine-protein kinase GSK3B, GSK-3 beta
检测方法:
ELISA
实验类型:
Sandwich
检测范围:
0.156-10ng/mL
灵敏度:
0.055 ng/mL
特异性:
Natural and recombinant human Glycogen synthase kinase-3 beta
样品类型:
Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
样品数据:
登录.
实验步骤:
研究领域:
Cancer
精密度
批内差:已知浓度的3个样本在一个板子内重复检测20次,以评估批内精密度。
批内 CV: ≤5.5%
批间差:已知浓度的3个样本在不同的板子上重复测定5次,以评估测定批间精密度。
批间 CV: ≤7.9%
批内 CV: ≤5.5%
批间差:已知浓度的3个样本在不同的板子上重复测定5次,以评估测定批间精密度。
批间 CV: ≤7.9%
回收率
回收率:低、中和高浓度的分析物被掺入到血清或者血浆样本中,进行回收实验测定。
Sample Type |
Average(%) |
Recovery Range(%) |
Serum |
93 |
87-99 |
Plasma |
95 |
89-101 |
线性
线性:给定样本通过梯度稀释,每次稀释的测量值与理论值的比值。
Sample |
1:2 |
1:4 |
1:8 |
1:16 |
serum(n=5) |
90-100% |
115-115% |
105-115% |
105-114% |
EDTA plasma(n=5) |
101-111% |
88-98% |
93-102% |
109-119% |
heparin plasma(n=5) |
102-114%
|
89-99% |
107-120% |
95-104% |
通用注释
亚单元:
Monomer. Interacts with ARRB2, DISC1 and ZBED3 (By similarity). Interacts with CABYR, MMP2, MUC1, NIN and PRUNE Interacts with AXIN1; the interaction mediates hyperphosphorylation of CTNNB1 leading to its ubiquitination and destruction. Interacts with and phosphorylates SNAI1. Interacts with DNM1L (via a C-terminal domain). Found in a complex composed of MACF1, APC, AXIN1, CTNNB1 and GSK3B (By similarity). Interacts with SGK3. Interacts with DAB2IP (via C2 domain); the interaction stimulates GSK3B kinase activation. Interacts (via C2 domain) with PPP2CA. Interacts with the CLOCK-ARNTL/BMAL1 heterodimer. Interacts with the ARNTL/BMAL1. Interacts with CTNND2 (PubMed:19706605). Interacts with NCYM (PubMed:24391509).
功能:
Constitutively active protein kinase that acts as a negative regulator in the hormonal control of glucose homeostasis, Wnt signaling and regulation of transcription factors and microtubules, by phosphorylating and inactivating glycogen synthase (GYS1 or GYS2), EIF2B, CTNNB1/beta-catenin, APC, AXIN1, DPYSL2/CRMP2, JUN, NFATC1/NFATC, MAPT/TAU and MACF1. Requires primed phosphorylation of the majority of its substrates. In skeletal muscle, contributes to insulin regulation of glycogen synthesis by phosphorylating and inhibiting GYS1 activity and hence glycogen synthesis. May also mediate the development of insulin resistance by regulating activation of transcription factors. Regulates protein synthesis by controlling the activity of initiation factor 2B (EIF2BE/EIF2B5) in the same manner as glycogen synthase. In Wnt signaling, GSK3B forms a multimeric complex with APC, AXIN1 and CTNNB1/beta-catenin and phosphorylates the N-terminus of CTNNB1 leading to its degradation mediated by ubiquitin/proteasomes. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Phosphorylates NFATC1/NFATC on conserved serine residues promoting NFATC1/NFATC nuclear export, shutting off NFATC1/NFATC gene regulation, and thereby opposing the action of calcineurin. Phosphorylates MAPT/TAU on 'Thr-548', decreasing significantly MAPT/TAU ability to bind and stabilize microtubules. MAPT/TAU is the principal component of neurofibrillary tangles in Alzheimer disease. Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. Phosphorylates MACF1, inhibiting its binding to microtubules which is critical for its role in bulge stem cell migration and skin wound repair. Probably regulates NF-kappa-B (NFKB1) at the transcriptional level and is required for the NF-kappa-B-mediated anti-apoptotic response to TNF-alpha (TNF/TNFA). Negatively regulates replication in pancreatic beta-cells, resulting in apoptosis, loss of beta-cells and diabetes. Through phosphorylation of the anti-apoptotic protein MCL1, may control cell apoptosis in response to growth factors deprivation. Phosphorylates MUC1 in breast cancer cells, decreasing the interaction of MUC1 with CTNNB1/beta-catenin. Is necessary for the establishment of neuronal polarity and axon outgrowth. Phosphorylates MARK2, leading to inhibit its activity. Phosphorylates SIK1 at 'Thr-182', leading to sustain its activity. Phosphorylates ZC3HAV1 which enhances its antiviral activity. Phosphorylates SNAI1, leading to its BTRC-triggered ubiquitination and proteasomal degradation. Phosphorylates SFPQ at 'Thr-687' upon T-cell activation. Phosphorylates NR1D1 st 'Ser-55' and 'Ser-59' and stabilizes it by protecting it from proteasomal degradation. Regulates the circadian clock via phosphorylation of the major clock components including ARNTL/BMAL1, CLOCK and PER2. Phosphorylates CLOCK AT 'Ser-427' and targets it for proteasomal degradation. Phosphorylates ARNTL/BMAL1 at 'Ser-17' and 'Ser-21' and primes it for ubiquitination and proteasomal degradation. Phosphorylates OGT at 'Ser-3' or 'Ser-4' which positively regulates its activity. Phosphorylates MYCN in neuroblastoma cells which may promote its degradation (PubMed:24391509).
亚细胞位置:
Cytoplasm
Nucleus
Cell membrane
The phosphorylated form shows localization to cytoplasm and cell membrane. The MEMO1-RHOA-DIAPH1 signaling pathway controls localization of the phosphorylated form to the cell membrane.
[1].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒可以做多少个样本?
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒分为2种规格,96孔和48孔。96孔的试剂盒,标曲和样本都做复孔的话,可以检测40个样本。96孔的试剂盒,标曲和样本都不做复孔的话,可以检测88个样本。
[2].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒使用视频?
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒实验操作视频在以下网址中,对每一步的实验步骤都做了演示,方便实验员能更好地理解ELISA实验的过程。
https://www.eiaab.com.cn/lesson-tech/805.html
https://www.eiaab.com.cn/lesson-tech/805.html
[3].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒是放在-20℃冰箱保存吗?
EIAab的人糖原合成酶激酶3β(GSK3B)ELISA试剂盒,洗涤液、底物、终止液保存于4℃,其余试剂-20℃冰箱保存。
[4].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒原理?
双抗体夹心法:用纯化的抗体包被微孔板,制成固相抗体,往包被有固相抗体的微孔中依次加入标准品或受检样本、生物素化抗体、HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
竞争法:用纯化的抗体包被微孔板,制成固相抗体,往包被有固相抗体的微孔中依次加入标准品或受检样本和生物素标记的目标分析物,受检标本中抗原与生物素标记抗原竞争结合有限的抗体。再加入HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
竞争法:用纯化的抗体包被微孔板,制成固相抗体,往包被有固相抗体的微孔中依次加入标准品或受检样本和生物素标记的目标分析物,受检标本中抗原与生物素标记抗原竞争结合有限的抗体。再加入HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
[5].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒中需要使用的样品量是多少?
夹心法100μL/孔,竞争法50μL/孔。如样本浓度过高时,应对样本进行稀释,以使稀释后的样本符合试剂盒的检测范围,计算时再乘以相应的稀释倍数。
[6].
如何分析人糖原合成酶激酶3β(GSK3B)ELISA试剂盒数据?
建议标准曲线,并计算样本浓度。对于elisa的曲线拟合,一般建议采用4参数曲线拟合,4参数曲线拟合通常更适合免疫分析。推荐使用专业软件进行曲线拟合,例如curve expert 1.3。根据样本的OD值由标曲查出相应的浓度,再乘以稀释倍数;或用标准物的浓度与OD值计算出标曲的回归方程式,将样本的OD值代入方程式,计算出样本浓度,再乘以稀释倍数,即为样本的实际浓度。以下链接是curve expert 1.3软件拟合曲线的方法。
https://www.eiaab.com.cn/news/502/
https://www.eiaab.com.cn/news/502/
[7].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒中是否包含人和动物的副产物,是否包含感染的或者传染性原料如HIV等?
除了抗体和稀释液中的BSA,不含其它人和动物的副产物,也不含感染材料。
[8].
收集人糖原合成酶激酶3β(GSK3B)ELISA试剂盒血浆样本,用什么作为抗凝剂?
一般建议用EDTA和肝素作为抗凝剂。
[9].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒酶标板可以拆成几部分?拆的时候是否需要避光,无菌?
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒酶标板是8×12孔条,可拆卸,板子可以拆成12条,注意避免孔污染,不需要避光和无菌。暂时不用的板子,放回原来装的袋子里,密封保存。
[10].
人糖原合成酶激酶3β(GSK3B)ELISA试剂盒样本如何保存?
尽量检测新鲜样本。若无新鲜样本,则4℃保存1周,-20℃保存1个月,-80℃保存2个月。
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