抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Human, Mouse, Rat, Pig, Horse
产品类型 一抗 磷酸化抗体
研究领域 肿瘤 神经生物学 信号转导 细胞凋亡 新陈代谢
蛋白分子量 predicted molecular weight: 22kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH conjugated Synthesised phosphopeptide derived from mouse BAD around the phosphorylation site of Ser112
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 WB=1:100-500 ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=1:100-500 ICC=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.
磷酸化相关死亡促进因子抗体产品介绍 Bad is a member of the Bcl2 family and acts to promote apoptosis by forming heterodimers with the survival proteins Bcl2 and BclxL, thus preventing them from binding with BAX. Bad is found on the outer mitochondrial membrane and, once phosphorylated in response to growth stimuli, translocates to the cytoplasm. The phosphorylation status of Bad represents a key checkpoint for death or cell survival. JNK-induced phosphorylation of BAD serine 128 promotes the apoptotic role of Bad by opposing the inhibitory effect of growth factor on Bad-mediated apoptosis. Cdc2-induced phosphorylation of Bad serine 128 has an inhibitory effect on its interaction with 14-3-3 proteins. The latter interaction is critical for Bad phosphorylation at serine 155, a site within the BH3 domain that leads to the release of BclxL and the promotion of cell survival. Alternative splicing of this gene results in two transcript variants which encode the same isoform.
Function : Promotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2. Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.
Subunit : Forms heterodimers with the anti-apoptotic proteins, Bcl-X(L), Bcl-2 and Bcl-W. Also binds protein S100A10. The Ser-75/Ser-99 phosphorylated form binds 14-3-3 proteins. Interacts with AKT1 and PIM3.
Subcellular Location : Mitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm.
Tissue Specificity : Expressed in a wide variety of tissues.
Post-translational modifications : Phosphorylated on one or more of Ser-75, Ser-99, Ser-118 andSer-134 in response to survival stimuli, which blocks itspro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75promotes heterodimerization with 14-3-3 proteins. This interactionthen facilitates the phosphorylation at Ser-118, a site within theBH3 motif, leading to the release of Bcl-X(L) and the promotion ofcell survival. Ser-99 is the major site of AKT/PKB phosphorylation, Ser-118 the major site of protein kinase A (CAPK) phosphorylation. Phosphorylation at Ser-99 by PKB/AKT1 is almost completely blockedby the apoptotic C-terminus cleavage product of PKN2 generated bycaspases-3 activity during apoptosis.
Methylation at Arg-94 and Arg-96 by PRMT1 inhibits Akt-mediated phosphorylation at Ser-99.
Similarity : Belongs to the Bcl-2 family.
Database links : UniProtKB/Swiss-Prot: Q92934.3
BAD是BCL2/BAX、BCL-XL/BAX异二聚体的负调节基因。BAD是BCL2/BCL-XL相关死亡促进因子,作为BCL2、 bCL-XL异二聚体伴分子而促进细胞凋亡。
有学者认为:BAD缺乏典型的羧基端跨膜结构,提示其并非一完整膜蛋白。与同BCL2作用相比,BAD与BCL-XL的结合更强,BAD以浓度依赖性方式替换BCL-XL/BAX、BCL2/BAX异二聚体中的BAX,使BAX游离而促进细胞凋亡。当一细胞系的所有细胞内异二聚体(BCL-XL/BAX和BCL2/BAX)的含量≥50%时,则细胞耐受凋亡;而当细胞内BAX同二聚体>80%时且在适当信号诱导下则细胞出现凋亡。这表明BAD通过调节BAX同二聚体与异二聚体量的比值而介导凋亡。
纯度:在实验的任何阶段,确定抗体溶液纯度的最简单方法是取一部分样本进行SDS-PAGE电泳。凝胶可用考马斯亮蓝染色(灵敏度为0.1—0.5ug/带)或银染(灵敏度1~l0ug/带)。
定量:如果抗体还不纯,有一个快捷的定量方法,即通过SDS-PAGE电泳分离出轻、重链,然后和已知的标准染色带比较。如果需要分析许多样本,用免疫测定法对抗体定量较容易。如果抗体是经过纯化的,可通过测蛋白总量代替上述两种方法,有一简单的方法,即紫外吸收法。磷酸化相关死亡促进因子抗体的量可通过测280nm处的吸收值来测(10D大致相当于0.75mg/m1的纯化抗体)。
抗原结合活性:一般说来,纯化方法不会引起抗原结合活性的改变。用蛋白G或蛋白A树脂很少导致抗体活性丧失。然而,如果最终抗体产物的作用不如原来所预料的好,检测抗体纯化过程所丢失的活性就极为重要。用一系列滴定法比较纯化的抗体和其原材料的活性,以标定每一步中的总抗体量,这将有助于较好的估计通过纯化所丢失的活性。