Anti-ATG5 Polyclonal Antibody is a Rabbit antibody targeting ATG5. Anti-ATG5 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Horse,Rabbit)

1. Paraformaldehyde-fixed, paraffin embedded (human gastric); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (ATG5) Polyclonal Antibody, Unconjugated (TMAB-00163) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (Human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (ATG5) Polyclonal Antibody, Unconjugated (TMAB-00163) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (ATG5) Polyclonal Antibody, Unconjugated (TMAB-00163) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Blank control: A431. Primary Antibody (green line): Rabbit Anti-ATG5 antibody (TMAB-00163)
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 room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
5. Paraformaldehyde-fixed, paraffin embedded (Rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (ATG5/APG5L) Polyclonal Antibody, Unconjugated (TMAB-00163) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-rabbit IgG antibody for 90 minutes, and DAPI for nucleus staining.
6. Sample:
Lane 1: Mouse Cerebrum tissue lysates
Lane 2: Mouse Kidney tissue lysates
Lane 3: Rat Cerebrum tissue lysates
Primary: Anti-ATG5/APG5 (TMAB-00163) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 32 kDa
Observed band size: 47 kDa

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

Polyclonal

KLH conjugated synthetic peptide: human APG5L

Human

Metabolism,Phagocytosis,APG gene products,Signal Transduction,Autophagy,APG gene products,Associated Proteins,Phagocytosis,Ub-like Proteins,Apoptosis,Autophagy and mitophagy,APG gene products