Antibody Type: Rabbit Polyclonal

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

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

IgG

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

1. Tissue/cell: human rectal carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-PKM2 Polyclonal Antibody, Unconjugated (TMAB-01545) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, FITC conjugated used at 1:200 dilution for 40 minutes at 37°C.
2. Tissue/cell: human lung cancer tissue; 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-PKM2 Polyclonal Antibody, Unconjugated (TMAB-01545) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Sample:
Lane 1: Testis (Rat) Lysate at 40 μg
Lane 2: Prostate (Rat) Lysate at 40 μg
Lane 3: Kidney (Rat) Lysate at 40 μg
Primary: Anti-PKM2 (TMAB-01545) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 60 kDa
Observed band size: 61 kDa
4. Blank control: A549. Primary Antibody (green line): Rabbit Anti-PKM2 antibody (TMAB-01545)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
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.

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: 1ug/Test

Polyclonal

KLH conjugated synthetic peptide: human PKM2

Human

5315

Energy Metabolism,Energy Metabolism,Carbohydrate metabolism,Kinases/Phosphatases,Enzymes,Metabolism of carbohydrates