Anti-PUMA antibody [EP512Y] (ab33906) at 1/5000 dilution + K562 cell lysate.

Predicted band size: 21 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

ab33906 at a 1:250 dilution staining PUMA in Hela cells using Immunocytochemistry/Immunofluorescence.

ab33906 at a 1:100 dilution staining PUMA in human ovary carcinoma using Immunohistochemistry, Paraffin Embedded Tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab33906 at a 1:100 dilution staining PUMA in human breast carcinoma using Immunohistochemistry, Paraffin Embedded Tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Overlay histogram showing K562 cells stained with ab33906 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33906, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in K562 cells fixed with methanol (5 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.