Anti-Wilms tumor/WT1 Polyclonal Antibody is a Rabbit antibody targeting Wilms tumor/WT1. Anti-Wilms tumor/WT1 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Paraformaldehyde-fixed, paraffin embedded (Mouse testis); 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 (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (TMAB-01979) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nucleus staining.
2. Sample: TM4 Cell (Mouse) Lysate at 40 μg
Primary: Anti-Wilms Tumor Protein (TMAB-01979) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 48 kDa
3. Tissue/cell: human kidney 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-WT-1/Wilms Tumor Protein Polyclonal Antibody, Unconjugated (TMAB-01979) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Blank control: Molt-4. Primary Antibody (green line): Rabbit Anti-Wilms Tumor Protein antibody (TMAB-01979)
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 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at 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 (Mouse kidney); 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 (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (TMAB-01979) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Sample:
Lane 1: Mouse Testis tissue lysates
Lane 2: Mouse Ovary tissue lysates
Lane 3: Mouse Liver tissue lysates
Lane 4: Human HL-60 cell lysates
Primary: Anti-WT1 (TMAB-01979) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 50 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 WT1

Human

Tumor biomarkers,Kidney development,Other factors,Tumor Associated