The document is a technical support questionnaire for a western blot experiment testing a GPR48 antibody on human embryonic stem cells (hESCs) and a shLGR4 knockdown cell line. The experiment aimed to detect a band around 100 kDa but no signal was observed for the predicted size in hESCs or the positive control. Information provided includes experimental details on sample preparation, gel electrophoresis, transfer, blocking, antibody incubation and detection conditions.