WT1 Rabbit Polyclonal Antibody

• Catalog Number: orb1621
• Category: Antibodies
• Description: WT1 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Gallus, Porcine, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human WT1 (301-400/449aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 55 kDa
• Target: WT1
• UniProt ID: P19544
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: WT1_HUMAN; Wilms tumor protein; WT33; WT1 transcri
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: Molt-4. Primary Antibody (green line): Rabbit Anti-Wilms Tumor Protein antibody (orb1621), 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. Acquisition of 20000 events was performed.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (orb1621) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (orb1621) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) for 90 minutes, and DAPI for nuclei staining.

Paraformaldehyde-fixed, paraffin embedded (Rat testis), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (orb1621) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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 (orb1621) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 55 kDa, Observed band size: 50 kDa.

Sample: TM4 Cell (Mouse) Lysate at 40 ug, Primary: Anti-Wilms Tumor Protein (orb1621) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 55 kD, Observed band size: 48 kD.