Anti-Cyclin E2 antibody [E142] (ab32103) at 1/500 dilution + HeLa cell lysate

Predicted band size: 45 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling Cyclin E2 with purified ab32103 at 1/2000. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

Immunofluorescent analysis of Cyclin E2 expression in HeLa cell culture using 1/100 ab32103.

Immunohistochemical analysis of Cyclin E2 expression in paraffin embedded human breast carcinoma using 1/50 ab32103.

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

Overlay histogram showing HeLa cells stained with ab32103 (red line). The cells were fixed with 80% methanol (5 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 (ab32103, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

ab32103 staining Cyclin E2 in the Hela cell line from Human cervix by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. Ab150081 (1/200) was used as the secondary antibody.

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