The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000 - 1/10000. Detects a band of approximately 146 kDa (predicted molecular weight: 146 kDa).
Use at 2-5 µg/mg of lysate.
Binds to the ubiquitin moieties of preformed conjugates and catalyzes ubiquitin chain assembly in conjunction with E1, E2, and E3.
Highest expression in ovary, testis, heart and skeletal muscle. Expression is low in colon, thymus and peripheral blood leukocytes. Almost undetectable in lung and spleen.
Belongs to the ubiquitin conjugation factor E4 family. Contains 1 U-box domain.
Proteolytically cleaved by caspases during apoptosis. Cleaved efficiently at Asp-123 by caspase-6 and granzyme B. Cleaved with approximately 10-fold less efficiency at Asp-109 by caspase-3 and caspase-7.
All lanes : Anti-UBE4B antibody (ab72497) at 0.04 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 Lane 5 : NIH3T3 whole cell lysate at 50 µg
Predicted band size : 146 kDa Observed band size : 146 kDa Additional bands at : 117 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds
Immunoprecipitation - UBE4B antibody (ab72497)
Detection of UBE4B by Western Blot of Immunprecipitate.
ab72497 at 0.1µg/ml staining UBE4B in HeLa whole cell lysate immunoprecipitated using ab72497 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection: Chemiluminescence with exposure time of 3 minutes.