抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Mouse, Rat, Dog, Rabbit
产品类型 一抗 磷酸化抗体
研究领域 肿瘤 细胞生物 神经生物学 信号转导 细胞凋亡
蛋白分子量 predicted molecular weight: 26kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH conjugated Synthesised phosphopeptide derived from rat Bcl-2 around the phosphorylation site of Ser70
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 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Bcl-2抗体产品介绍 BCL2 is an integral outer mitochondrial membrane protein that blocks the apoptotic death of some cells such as lymphocytes. Constitutive expression of BCL2, such as in the case of translocation of BCL2 to Ig heavy chain locus, is thought to be the cause of follicular lymphoma. Two transcript variants (alpha and beta) produced by alternate splicing, differ in their C-terminal ends. BCL2 suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. It regulates cell death by controlling the mitochondrial membrane permeability. It appears to function in a feedback loop system with caspases. BCL2 inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF1). It can form homodimers, and heterodimers with BAX, BAD, BAK and BclX(L). Heterodimerization with BAX requires intact BH1 and BH2 domains, and is necessary for anti-apoptotic activity. Also interacts with APAF1, RAF1, TP53BP2, BBC3, BCL2L1 and BNIPL.
Function : Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1).
Subunit : Forms homodimers, and heterodimers with BAX, BAD, BAK and Bcl-X(L). Heterodimerization with BAX requires intact BH1 and BH2 motifs, and is necessary for anti-apoptotic activity. Interacts with EI24 (By similarity). Also interacts with APAF1, BBC3, BCL2L1, BNIPL, MRPL41 and TP53BP2. Binding to FKBP8 seems to target BCL2 to the mitochondria and probably interferes with the binding of BCL2 to its targets. Interacts with BAG1 in an ATP-dependent manner. Interacts with RAF1 (the 'Ser-338' and 'Ser-339' phosphorylated form). Interacts (via the BH4 domain) with EGLN3; the interaction prevents the formation of the BAX-BCL2 complex and inhibits the anti-apoptotic activity of BCL2. Interacts with G0S2; this interaction also prevents the formation of the anti-apoptotic BAX-BCL2 complex.
Subcellular Location : Mitochondrion outer membrane; Single-pass membrane protein. Nucleus membrane; Single-pass membrane protein. Endoplasmic reticulum membrane; Single-pass membrane protein.
Tissue Specificity : Expressed in a variety of tissues.
Post-translational modifications : Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
Monoubiquitinated by PARK2, leading to increase its stability.
DISEASE : Note=A chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions.
Similarity : Belongs to the Bcl-2 family.
Database links : UniProtKB/Swiss-Prot: P10415.2 (humna)
UniProtKB/Swiss-Prot: P49950.2 (rat)
UniProtKB/Swiss-Prot: P10417.3(mouse)
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Bcl-2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。
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