Rab35, Abcam, ab288567

Our team performed a comprehensive review of the scientific literature to identify studies using antibodies against human Rab35, followed by extraction and evaluation of available data according to pAbmAbs rating criteria.

YCharOS conducted a systematic head-to-head comparison of 6 commercially available antibodies against Rab35 across multiple applications.

Antibodies were tested by WB using lysates (20 µg protein) from HAP1 WT cells and HAP1 Rab35 KO cells. These cell lines were also used for IF. In addition, antibodies were evaluated by IP using HAP1 cell lysates (0.5 mg protein). All experiments were performed according to YCharOS standard protocols (see reference).

In WB (1:1000 antibody dilution), ab288567 detected a clear band at the predicted size (~23 kDa) in WT HAP1 cell lysate, with no corresponding band in Rab35 KO lysate. Additionally, several weaker bands of higher molecular weight were observed in both cell lysates.

In IP, the antibody (2 µg) did not enrich Rab35 upon precipitation. Rab35 levels in the starting material, unbound fraction, and precipitate were assessed by WB using the Rab35-specific antibody ab300116 (Abcam)

In IF, the antibody (1:500 dilution) produced a Rab35-specific staining distributed throughout HAP1 WT cells, with weak background signal in Rab35 KO cells.

We highly encourage other scientists to share their observations.