| Rabbit Anti-Phospho-Bcl2 (Thr69) antibody |
| 产品应用（已验证） |
WB,IHC,FCM |
| 产品应用（可尝试） |
ICC,IF,ELISA |
| 推荐稀释比例 |
WB=1:500-2000,Elisa=1:5000-10000,IHC-P=1:100-500,IHC-F=1:100-500,Flow Cyt=1μg/Test,IF=1:100-500,ICC=1:100, |
| 研究领域 |
肿瘤,细胞生物,神经生物学,信号转导,细胞凋亡,新陈代谢,线抗体, |
| 标签 |
Array |
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Blank control:HL-60.
Primary Antibody (green line): Rabbit Anti-Phospho-Bcl2 (Thr69) antibody (bs-12578R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1%PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Blank control(blue):K562 (fixed with 2% paraformaldehyde for 10 min at 37℃).
Primary Antibody:Rabbit Anti-Phospho-Bcl2 (Thr69) antibody (bs-12578R,Green); Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions;
Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
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Paraformaldehyde-fixed, paraffin embedded (rat stomach tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (P-Bcl2 (Thr69)) Polyclonal Antibody, Unconjugated (bs-12578R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
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Tissue/cell: rat lung tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-Bcl2 (Thr69) Polyclonal Antibody, Unconjugated(bs-12578R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Sample:
Spleen (Mouse) Lysate at 40 ug
LOVO Cell (Human) Lysate at 40 ug
HL60 Cell (Human) Lysate at 40 ug
U937 Cell (Human) Lysate at 40 ug
MCF-7 Cell (Human) Lysate at 40 ug
Hela Cell (Human) Lysate at 40 ug
Primary: Anti- Phospho-Bcl2 (Thr69) (bs-12578R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 26 kD
RRID：RRID
产品名称：Rabbit Anti-Phospho-Bcl2 (Thr69) antibody
别名： Bcl2 (Phospho-Thr69); Bcl2 (Phospho-T69); Bcl2 (phospho T69); p-Bcl2 (phospho T69); Apoptosis regulator Bcl 2; Apoptosis regulator Bcl2; AW986256; B cell CLL/lymphoma 2; B cell leukemia/lymphoma 2; B cell lymphoma 2; Bcl 2; Bcl-2; Bcl2; BCL2 protein; C430
中文名称：磷酸化Bcl-2抗体
英文名称：Rabbit Anti-Phospho-Bcl2 (Thr69) antibody
抗体来源： Rabbit
克隆类型：多克隆
细胞定位：细胞核,细胞浆,细胞膜
性 状：Liquid
亚 型：IgG
纯化方法：affinity purified by Protein A
保存条件：Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
免 疫 原：KLH conjugated Synthesised phosphopeptide derived from human Bcl-2 around the phosphorylation site of Thr69
抗原表位：AR(p-T)SP
SWISS：P49950
Gene ID ：596
Human Gene ID：596
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. In
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,
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 grow
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 attr
Similarity：Belongs to the Bcl-2 family.
Important Note：This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
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