R & D Systems
DGAT2 Antibody
AF7256
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Aim of the study was to test the effect of a growth factor treatment on human primary cell line on DGAT2 protein levels. For this, cells starved for 12 hours were treated with growth factor, lysed and protein extracts were isolated. An equal amount of proteins was resolved in denaturing gel and immunoblotted with DGAT2 antibody obtained from R & D systems. Although antibody was also available with other suppliers, the selection was solely based on the information’s indicated in antibody flyer. Low range protein marker from Bio-Rad was used.
Western Blot
Human primary cell whole cell lysate
Dilution (1:1500) in 5% BSA prepared in 1x TBST, overnight incubation at 4 degree Celsius
skimmed milk (5%) prepared in 1x TBST
Goat anti-Sheep IgG HRP, dilution (1:4000) in 5% skimmed milk prepared in 1x TBST, 1hr at RT
None
ECL detection in X-Ray film
Although, the antibody has detected a specific band of AMPKα at the correct molecular weight (compared with low range protein marker, Bio-Rad# 161-0304) few non-specific bands were also detected. The difference of antibody non-specificity between flyer and the present experiment is maybe due different cell types and experimental conditions.
Non-specific bands were also observed
Detected a specific band of DGAT2
The antibody also cross-reacted with other nonspecific epitopes
Best antibody for the DGAT2 detection in human cells