Anti-BAK1 Polyclonal Antibody is a Rabbit antibody targeting BAK1. Anti-BAK1 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse (predicted:Rat)

1. Tissue/cell: human colon carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Bak Polyclonal Antibody, Unconjugated (TMAB-00183) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Sample: Lung (Mouse) Lysate at 30 μg
Primary: Anti-Bak (TMAB-00183) at 1:300 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit Igg (bse-0295G) at 1: 5000 dilution;
Predicted band size: 23 kDa
Observed band size: 23 kDa
3. Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Bak Polyclonal Antibody, Unconjugated (TMAB-00183) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Blank control: Hela (blue). Primary Antibody: Rabbit Anti-Bak antibody, Dilution: 1 μg 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-Pe (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Antibody (TMAB-00183, 1 μg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of TMAB-00183 at 1/200 dilution for 30 min on ice.
5. Blank control: THP-1. Primary Antibody (green line): Rabbit Anti-Bak antibody (TMAB-00183)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST methanol 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.

WB,IHC-P,IHC-Fr,IF,FCM

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

Polyclonal

KLH conjugated synthetic peptide: mouse Bak

Mouse

Bcl2 Family