Immunocytochemistry/Immunofluorescence analysis of C6 (rat glioma) labelling MAP1LC3A with purified ab52768 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

Overlay histogram showing SHSY-5Y cells stained with ab52768 (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 (ab52768, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Anti-MAP1LC3A antibody [EP1983Y] - Autophagosome Marker (ab52768) at 1/10000 dilution + Hela cell lysate at 10 µg

Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution

Predicted band size: 14 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?

Immunocytochemistry/Immunofluorescence analysis of human keratinocytes (pannel A) and VZV-infected melanoma cells (panel E) labelling MAP1LC3A with ab52768.

Immunohistochemical staining of MAP1LC3A in paraffin embedded human breast carcinoma using ab52768 at a 1/100 dilution.

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