The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 125 kDa (predicted molecular weight: 113 kDa).
Use a concentration of 5 µg/ml.
CDCA2 is a regulator of chromosome structure during mitosis. CDCA2 forms an essential complex with protein phosphatase PP1-gamma subunit 1 (PPP1CC) and is required for the recruitment of PP1 to chromatin. Knockdown of CDCA2 by RNAi causes large-scale cell death by apoptosis, as does overexpression of a dominant-negative mutant of the protein.
ICC/IF image of ab45129 stained HepG2 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 (ab45129, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.