Overlay histogram showing HL60 cells stained with ab55528 (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. The cells were then incubated with the antibody (ab55528, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HL60 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunoprecipitation - Anti-MDH1 antibody [2B11-B7] (ab55528)This image is courtesy of an anonymous Abreview
Detection of MDH1 by Immunoprecipitation. ab55528 used at 5µl/mg for 500µg of Mouse whole cell lysate from Hippocampal neurol cells for IP using Dynabeads for 16 hours at 4 degrees (in PBST). Immunoprecipitated MAD2 was blotted with another antibody HRP-congugated Goat anti-mouse Polyclonal at 1/5000. Detection: Chemiluminescence with an exposure time of 1 minute. Band size 36kDa.