bs-5220R [Primary Antibody]
Bcl-2(Thr129) Polyclonal Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: Bcl-2Thr129

Modification Site: Thr129

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 24224

Source: KLH conjugated synthetic phosphopeptide derived from rat Bcl-2 around the phosphorylation site of Thr129

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Background:

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.

Size: 100ul

Concentration: 1ug/ul

Applications: WB(1:300-5000)
ELISA(1:500-1000)
FCM(1:20-100)
IHC-P(1:200-400)
IHC-F(1:100-500)
IF(IHC-P)(1:50-200)
IF(IHC-F)(1:50-200)
IF(ICC)(1:50-200)

Predicted Molecular Weight: 26


Cross Reactive Species: Human
Mouse
Rat
Others

Predicted Cross Reactive Species: Dog
Cow
Sheep
Pig
Horse

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Antigen: bs-5220P, 0.2ug/100ul \nPrimary: Antiserum, 1:500, 1:1000, 1:2000, 1:4000, 1:8000, 1:16000, 1:32000; \nSecondary: HRP conjugated Goat Anti-Rabbit IgG(bs-0295G-HRP) at 1: 5000; \nTMB staining; Read the data in Microplate Reader by 450nm\n


Mouse heart lysates probed with Phospho-Bcl-2 (Thr129) Polyclonal Antibody, Unconjugated (bs-5220R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.


HL-60 cell line lysates probed with Bcl-2 (Thr129) Polyclonal Antibody, Unconjugated (bs-5220R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.


Paraformaldehyde-fixed, paraffin embedded Mouse brain; 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 Bcl-2 (Thr129) Polyclonal Antibody, Unconjugated (bs-5220R) at 1:400 overnight at 4°C, DAB staining.


HL-60 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Phospho-Bcl-2 (Thr129) Polyclonal Antibody(bs-5220R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


Paraformaldehyde-fixed, paraffin embedded Rat spleen; 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 Bcl-2(Thr129) Polyclonal Antibody, Unconjugated (bs-5220R) at 1:200 overnight at 4°C, DAB staining.