GAPDH Antibody

• Catalog Number: orb1925320
• Category: Antibodies
• Description: Purified Mouse Monoclonal Antibody (Mab)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 1653CT614.35.51
• Tested applications: FC, IHC-P, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG1,k
• Dilution range: WB: 1:8000, IHC-P: 1:25, IHC-P: 1:25, FC: 1:25
• Form/Appearance: Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein G column, followed by dialysis against PBS.
• Conjugation: Unconjugated
• MW: 36053 Da
• Target: This GAPDH antibody is generated from a mouse immunized with a recombinant protein between 43-335 amino acids from human GAPDH.
• UniProt ID: P04406
• 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
• Alternative names: Glyceraldehyde-3-phosphate dehydrogenase, GAPDH, 1
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

All lanes: Anti-GAPDH Antibody at 1:8000 dilution. Lane 1: Hela whole cell lysate. Lane 2: Jurkat whole cell lysate. Lane 3: A549 whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 36 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Staining GAPDH in human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

Staining GAPDH in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

Overlay histogram showing Hela cells stained (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at 37°C. Isotype control antibody (blue line) was mouse IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 events was performed.