龙虎山
bs-0032R [Primary Antibody]
Rabbit  Anti-Bcl-2 Polyclonal Antibody
www.egfqj.club
[email protected]
[email protected]
400-901-9800
DATASHEET

Host: Rabbit

Target Protein: Bcl-2

IR: Immunogen Range:101-160/236

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 596

Swiss Prot: P49950

Source: KLH conjugated synthetic peptide derived from human Bcl-2:101-160/236 

Purification: affinity purified by Protein A

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. 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.

Background: The Bcl-2 gene was isolated at the chromosomal breakpoint of t(14;18)-bearing follicular B cell lymphomas(1,2).Bcl-2 blocks cell death following a variety of stimuli and confers a death-sparing effect to certain hematopoietic cell lines following growth factor withdrawal (3,5).Bcl-2 appears to function in several subcellular locations yet lacks any known motifs that would confer insight into its mechanism of action (6,7).A more recently identified protein,designated Bax p21(i.e., Bcl-associated X protein ),has extensive amino acid homology with Bcl-2 and both homodimerizes and forms heterodimers with Bcl-2(8). Overexpression of Bax accelerates apoptotic death induced by cytokine deprivation in an IL-3 dependent cell line and Bax also counters the death repressor activty of Bcl-2(8).

Size: 100ul

Concentration: 1mg/ml

Applications: WB(1:500-2000)
ELISA(1:500-1000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(3ug/test)
IF(1:100-500)

Cross Reactive Species: Human
Chicken
Dog
Pig
Cow
Horse
Rabbit
Guinea Pig
.

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

VALIDATION IMAGES
Protein: Spleen(Mouse)lysate 30ug;
Primary: Anti-Bcl-2(bs-0032R) at 1:300;
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Predicted band size : 26kD
Observed band size : 26kD
Protein: Recombinant protein lysate 100ng;
Primary: Anti-Bcl-2(bs-0032R) at 1:200;
Secondary: HRP conjugated Goat Anti-Rabbit IgG(bs-0295G-HRP) at 1: 5000;
Predicted band size : 26kD
Observed band size : 26kD
Protein: Brain(Mouse)lysate 30ug;
Primary: Anti-Bcl-2(bs-0032R) at 1:200;
Secondary: HRP conjugated Goat Anti-Rabbit IgG(bs-0295G-HRP) at 1: 5000;
Predicted band size : 26kD
Observed band size : 26kD
Sample:
Bcl-2 overexpression E.coli lysate (whole cell) Lysate at 40 ug
Recombinant protein Lysate at 40 ug
Primary: Anti-Bcl-2 (bs-0032R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 25 kD
Sample:Spleen (Mouse) Lysate at 40 ug
Primary: Anti-Bcl-2 (bs-0032R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 26 kD
Sample:
MCF-7(Human) Cell Lysate at 30 ug
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-Bcl-2 (bs-0032R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 30 kD
Sample:
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti-Bcl-2 (bs-0032R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 26 kD
Sample:
Spleen (Mouse) Lysate at 40 ug
RAW264.7 Cell (Mouse) Lysate at 40 ug
Primary: Anti-Bcl-2 (bs-0032R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 26 kD
Paraformaldehyde-fixed, paraffin embedded (human pancreatic cancer); 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) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human pancreatic cancer); 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) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human pancreatic cancer); 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) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat spinal cord); 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) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat 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) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat ovary 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 (Bcl-2) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat thyroid gland); 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 (B cell lymphoma 2; Bcl-2) Polyclonal Antibody, Unconjugated (bs-0032R) at 1:200 overnight at 4°C, followed by a conjugated secondary antibody (bs-0295G-cy3) for 90 minutes and DAPI for nuclei staining.
Blank control: Jurkat.
Primary Antibody (green line): Rabbit Anti-Bcl-2 antibody (bs-0032R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. 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. Acquisition of 20,000 events was performed.
 
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