Abcam
Anti-Ubiquitin (linkage-specific K48) antibody
ab140601
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This antibody was used to detect polyubiquitin chains formed by Lysine-48 (K48) residue linkages via western blot. K48 ubiquitin linkages are involved in ubiquitin proteasome system protein degradation. This product was used in conjunction with a PAN ubiquitin antibody and a positive control, Mg132. It was selected due to detection observed in agreement with PAN ubiquitination, and a significant increase when used in the presence of a positive control.
Western Blot
H4IIE Whole cell lysates
1:1000, 4C, overnight with gentle shaking in 1:3 Odyssey Blocking buffer (Licor):TBST (0.1% tween20)
1:3 Odyssey Blocking buffer:TBS
1:15,000 goat-anti-rabbit (Licor IRDye800CW) at room temperature with gentle shaking in 1:3 Odyssey Blocking buffer (Licor):TBST (0.1% tween20, 0.02% SDS)
N/A
Odyssey Clx
This antibody required minimal optimization. It has produced consistent results and seems to work well in order to distinguish the lysine-specific residue that is being ubiquitinated.
Loaded 25ug of protein
Great for detecting lysine-specific protein ubiquitination
None
Great antibody for detecting K48 linkage-specific protein ubiquitination via western blot. Requires little optimization, and has produced consistent results.