3-磷酸甘油醛脱氢酶(兔来源免疫组化用抗体)英文名称 Anti-GAPDH
中文名称 3-磷酸甘油醛脱氢酶(兔来源免疫组化用抗体)
别 名 Glyceraldehyde-3-phosphate dehydrogenase.
浓 度 1mg/1ml
规 格 0.1ml/100μg 0.2ml/200μg
纯化的抗体可通过不同的途径获取,有些3-磷酸甘油醛脱氢酶(兔来源免疫组化用抗体)可通过下述方法制备或从商家购买。从商家购买的抗体,通常附有正确的储存方法。
1)工作液应在4℃下融化并存放,可能稳定达数月。
2)如果没有特殊原因而避免使用叠氮钠,亦可加入叠氮钠,浓度为0.02%。将纯化的抗体样本分装成合适的体积,于-20℃保存。
3)纯化的抗体溶液应以较高的浓度(如lmg/m1)在中性pH下保存。:常用的抗体储存浓度高达l0mg/ml。较低浓度的抗体冻存前应浓缩。所有标准的浓缩方法(如超滤法),皆可使用。还有一个简单的方法是用蛋白A或蛋白G亲和柱来浓缩溶液。如果纯化的抗体不是用于标记,可将它们以较低浓度储存于加有1%BSA的溶液中。
4)经纯化制备的抗体在常用的缓冲液中是稳定的。其DH应保持在中性左右。如果pH在7-8之间,即使保存多年,对抗体也无损害。多数情况下,盐浓度适于保持在0-150mmol/L之间,但在长期存放的抗体中,盐溶液浓度高达500mmol/L时,对3-磷酸甘油醛脱氢酶(兔来源免疫组化用抗体)能有损害。如果没有其他说明.律议用PBS或50mmol/LTris(DH8.0)溶液长期存放抗体。
抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Human, Mouse, Rat
产品类型 一抗
研究领域 细胞生物 免疫学
蛋白分子量 predicted molecular weight: 37kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from the middle of human GAPDH
亚 型 IgG
纯化方法 affinity purified by Protein A
储 存 液 0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide
产品应用 ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:100-500
(石蜡切片需做抗原修复)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
保存条件 Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
Important Note This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
3-磷酸甘油醛脱氢酶(兔来源免疫组化用抗体)产品介绍 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. It catalyzes an important energy-yielding step in carbohydrate metabolism, thereversible oxidative phosphorylation of glyceraldehyde-3-phosphatein the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). Recent evidence suggests that it also is involved in a number of cellular processes such as membrane fusion, phosphotransferase activity, DNA replication and repair, and nuclear RNA export (1). GAPDH has also been implicated in playing a role in different pathologies such as cancer progression, apoptosis, and neuronal diseases such as Alzheimer’s and Huntington’s disease (2). GAPDH is constitutively expressed at high levels in almost all tissues and cell lines making it ideal for use as a loading control marker in immunoblots.
Function : Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.
Subunit : Homotetramer. Interacts with TPPP; the interaction is direct. Interacts (when S-nitrosylated) with SIAH1; leading to nuclear translocation. Interacts with RILPL1/GOSPEL, leading to prevent the interaction between GAPDH and SIAH1 and prevent nuclear translocation. Interacts with EIF1AD, USP25, PRKCI and WARS.
Subcellular Location : Cytoplasm, cytosol. Nucleus. Cytoplasm, perinuclear region. Membrane. Note=Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal. Postnuclear and Perinuclear regions.
Post-translational modifications : S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated (Probable).
Sulfhydration at Cys-152 increases catalytic activity.
Similarity : Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
甘油醛-3-磷酸脱氢酶(Glyceraldehyde 3 phosphate dehydrogenase,GAPDH)是糖酵解(glycolysis)过程中的关键酶。除了在胞质中作为糖酵解的酶以外,有证据表明哺乳动物细胞中的GAPDH参与了多种胞内生化过程,包括膜融合(membrane fusion)、微管成束(microtubule bundling)、磷酸转移酶(phosphotransferase)激活、核内RNA出核、DNA复制与DNA修复。一些生理因素,诸如低氧(hypoxia)和尿糖(diabetes),可以增加GAPDH在特定细胞中的表达。GAPDH存在于几乎所有的组织中,以高水平持续表达。
GAPDH(甘油醛-3-磷酸脱氢酶)是参与糖酵解的一种关键酶,由4个30-40kDa的亚基组 GAPDH蛋白几乎在所有组织中都高水平表达,广泛用作Western blot蛋白质标准化的内参,是很好的内参抗体。
GAPDH 作为管家基因在同种细胞或者组织中的蛋白质表达量一般是恒定的。在实验中,可能存在总蛋白浓度测定不准确;或者蛋白质样品在电泳前上样时产生的样品间的操作误差;这些误差需要通过测定每个样品中实际转到膜上的GAPDH的含量来进行校正,所以一般的western实验都需要进行内参设置。具体校正的方法就是将每个样品测得的目的蛋白含量与本样品的GAPDH含量相除,得到每个样品目的蛋白的相对含量。然后才进行样品与样品之间的比较。