I want to measure ubiquitination following IP of my proteins of interest. Does anyone have experience with ubiquitin antibodies that work well in skeletal muscle tissue or L6 muscle cells?
Hello
E3 ubiquitin ligase that, together with UBE2N and UBE2V1, mediates the synthesis of 'Lys63'-linked polyubiquitin
Please , read articles in attachment , i hope help you
Good Luck
Anti-Ub P4D1 clone,info in the link
http://www.enzolifesciences.com/BML-PW0930/ubiquitin-mab-p4d1/
Thank you very much Dr Alkarim and Dr Pokrzywa
Dr Pokrzywa, which dilution and conditions have you used this antibody with? Yhank you
They are quite specific, but of course depends on the protein amount,success of your IP etc. We performed IP from C2C12 cells, detection with anti-Ub 1/5000 in 3 milk PBS-T, let say over night. Secondary 1/10000, 45min.
Check the conditions firstly on your input,before IP. For us 25uq of proteins in the lane was quite enough to see a polyubiquitylation patterns.
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