Cell Signaling Technology
Ubiquitin (P4D1) Mouse mAb
3936
Monoclonal
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This antibody detects ubiquitylated proteins in cell lysates with high sensitivity. I recommend using low % acryamide gel to probe high molecular weight proteins with poly-ubiquitin chains.
Western Blot
HeLa cell lyate
1:2000 dilution, 4C overnight.
5% non-fat milk
1:20000 dilution of anti-mouse IRDye-800
Washing with TBST X3, 5 min each
Odyssey
I confirmed this antibody using UBE1 inhibitor MLN7243. To resolve free ubiquitin, I would use high % acrylamide gel. I used NuPAGE-bis-tris gel 4-12% with MES buffer, which gave nice ubiquitylated substrate bands. However, the free ubiquitin band appeared as smear at the bottom of the blot.
N/A
Good quality
Nothing
Sensitive antibody for total ubiquitylated substrates.