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                          bs-0032R [Primary Antibody]
                          Rabbit  Anti-Bcl-2 Polyclonal Antibody
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                          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: 200ul

                          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
                          Mouse
                          Rat
                          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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