Novus Biologicals
Anti-53BP1 antibody
NB100-304
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This study was done to investigate the protein expression regulation of 53BP1 in cells. Therefore, immunoblotting of 53BP1 was a major readout in our study. We revealed that UbcH7 regulates the proteasome dependent degradation of 53BP1. This study was published in Proc Natl Acad Sci U S A. 2014 Nov 24. pii: 201408538.
Western Blot
Cultured human cell lines
1:1000 dilution in 1X TBST for overnight at 4 degrees
5% BSA
Anti-rabbit 1:7500 in 1X TBST with 3-5% dry milk at room temp for 1 hour
None
ECL
Overall, this antibody works well for detection of 53BP1 proteins from cultured human cells. The protein runs much slower than the predicted molecular size as reported (well above the 250 kDa marker). However, it appears that the quality of the blot varies over time and still there is no definite solution for that. This can be seen from the two sample results attached here. While in result 1, it only shows a clear band corresponding to 53BP1, in other times like in result 2, there are two extra bands running faster than the one in result 1.
Works for both Western blot and immunofluorescence.
Sometimes too much signal for the extra bands.
It is able to detect 53BP1 in both Western blot and immunofluorescence.