The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/200. Incubate for 30 minutes at RT. Antigen retrieval is recommended: boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.
1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionSpecific inhibition of calpain (calcium-dependent cysteine protease). Plays a key role in postmortem tenderization of meat and have been proposed to be involved in muscle protein degradation in living tissue.
Sequence similaritiesBelongs to the protease inhibitor I27 (calpastatin) family.
DomainHas four inhibitory domains.
Post-translational modificationsThe N-terminus is blocked.
Overlay histogram showing HeLa cells stained with ab101684 (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 (ab101684, 1/100 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) (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.