FunctionProbable E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. May be required for development of the head mesenchyme and neural tube closure.
PathwayProtein modification; protein ubiquitination.
ab101992 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab101992 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Western blot - HECTD1 antibody (ab101992)
All lanes : Anti-HECTD1 antibody (ab101992) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg
developed using the ECL technique
Predicted band size : 290 kDa
Exposure time : 3 minutes
Immunoprecipitation - HECTD1 antibody (ab101992)
ab101992 at 1 µg/ml staining HECTD1 in HeLa cell lysate immunoprecipitated using ab101992 at 3 µg/mg lysate (1 mg/IP; 20% of IP loaded/lane).
Detection: Chemiluminescence with exposure time of 3 minutes
References for Anti-HECTD1 antibody (ab101992)
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