ANAPC2 Antibody LS-C356339
LS-C356339 is an unconjugated rabbit polyclonal antibody to human ANAPC2 (aa712-746). Validated for IHC, Peptide-ELISA and WB.
ANAPC2 antibody a multi-subunit complex that regulates the turnover of many proteins, including mitotic regulators and cell fate determinants. Recently, mechanistic studies indicated that APC/C-Cdh1 controls neuronal development by targeting SnoN and Id2 for degradation [145, 146]. APC/C also regulates apoptosis through targeted degradation of pro-apoptotic proteins such as Bim and Moap [147]. These results suggest that the activity of APC/C is essential in cellular functions ranging from organ growth and patterning to regulating cell cycle progression and cell death.
ANAPC2 Antibody: Uses and Applications in Research
To establish a quantitative system for monitoring APC/C-dependent protein degradation in single cells, we monitored Venus-ANAPC4 and deGradFP fluorescence over time in tetracycline-inducible LMNA cells. We found that both mAID-nanobody and deGradFP efficiently targeted Venus-ANAPC4 for degradation within 3 h of auxin addition (Fig. 5a and Supplementary Movie 1). The degradation efficacy of the alternative nanobody was slightly less than that of the mAID-nanobody, likely because the additional time required for deGradFP expression and folding might have led to protein stabilization.
We further confirmed the degradation kinetics observed by live cell imaging using Western blot. Degradation of ANAPC4 by the mAID-nanobody occurred in the nucleus and cytoplasm with similar efficiency, but not in mitochondria. We also found that the deGradFP-targeted ANAPC4 was partially but not completely depleted after auxin withdrawal, and this effect was significantly enhanced when proteasome inhibition with MG132 was used (Supplementary Figure 3g). Judging by the remarkably low levels of ANAPC4, ANAPC3, ANAPC10 and ANAPC12 depleted by the mAID-nanobody, we concluded that the mAID-nanobody targets ANAPC2/ANAPC10 for SCFTIR1-mediated ubiquitylation/degradation in the APC/C complex.…