Anti-MCM2 antibody (ab4461)

  Western blot - MCM2 antibody (ab4461)

Predicted band size : 100 kDa


Sample: HeLa cell RIPA extract.

WB - 50 µg.

CoIP - 7 µg.

ab4461 incubated at 0.2 µg/ml for 1 hour for WB.

ab4461 incubated at 20 µg for CoIP.

Detection:

WB -ECL

CoIP - Coomassie Stain

  Immunocytochemistry/ Immunofluorescence-MCM2 antibody(ab4461)

ICC/IF image of ab4461 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4461, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-MCM2 antibody(ab4461)

Ab4461 staining MCM2 in Human placenta.
Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.