Rab6A, Cell Signaling Technology, 9625

Our team performed a comprehensive review of the scientific literature to identify studies using antibodies against human Rab6A, followed by extraction and evaluation of available data according to pAbmAbs rating criteria.

YCharOS conducted a systematic head-to-head comparison of 13 commercially available antibodies against Rab6A across multiple applications

Antibodies were tested by WB using lysates (40 µg protein) from HAP1 WT cells and HAP1 Rab6A KO cells, which may express a truncated lower molecular weight variant. These cell lines were also used for IF. In addition, antibodies were evaluated by IP using HAP1 cell lysates (1 mg protein). All experiments were performed according to YCharOS standard protocols (see reference).

In WB (1:100 antibody dilution), 9625 detects a weak band at the expected size (~24 kDa) in WT HAP1 cell lysate with no corresponding band in lysate from Rab6A KO cells. In addition several more intense background bands of higher molecular weight are detected in both lysates.

In IP, the antibody (2 µg) enriches Rab6A following precipitation, as detected by WB using the Rab6A-specific antibody NBP3-13701. However, the band in the IP fraction is only slightly more intense than in the starting material, and the target band remains detectable in the unbound fraction, indicating partial but incomplete immunoprecipitation.

In IF, the antibody (1:500 dilution) resulted in a clear staining of HAP1 WT with on background signal in Rab6A KO cells.

We highly encourage other scientists to share their observations.