Anti-Bad Polyclonal Antibody is a Rabbit antibody targeting Bad. Anti-Bad Polyclonal Antibody can be used in WB, IHC-P, IHC-Fr, IF, FCM.

IgG

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (rat stomach); 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 30 min; Antibody incubation with (Bad) Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Sample: Hela Cell lysate 30 ug;
Primary: Anti-Bad (TMAB-03008) at 1:300;
Secondary: HRP conjugated Goat -Anti-Rabbit IgG (Goat Anti-Rabbit IgG H&L-HRP) at 1:5000;
Predicted band size : 18 kD
Observed band size : 27 kD
3. Sample: Raji Cell lysate 30 ug;
Primary: Anti-Bad (TMAB-03008) at 1:300;
Secondary: HRP conjugated Goat -Anti-Rabbit IgG (Goat Anti-Rabbit IgG H&L-HRP) at 1:5000;
Predicted band size : 18 kD
Observed band size : 27 kD
4. 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 30 min; Antibody incubation with (Bad) Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Tissue/cell: Human kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Bad Polyclonal Antibody, Unconjugated (TMAB-03008) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
6. Blank control: A431(Black).
Primary Antibody (green line): Rabbit Anti-Bad antibody (TMAB-03008)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min 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 -20℃. 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.
7. Blank control (Black line): Jurkat (Black).
Primary Antibody (green line): Rabbit Anti-Bad antibody (TMAB-03008)
Dilution: 3μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% Pspecific protein-protein interactions for 15 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.
8. Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Bad Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L-HRP and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Bad Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L-HRP and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Bad Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L-HRP and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Bad Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L-HRP and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Bad Polyclonal Antibody, Unconjugated (TMAB-03008) at 1:200 overnight at 4°C, followed by conjugation to the Goat Anti-Rabbit IgG H&L-HRP and DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 3μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Bad

Human

BAD

Bcl2-associated agonist of cell death

Promotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2 (By similarity). Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.