Rab35, Abcam, ab300116

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), ab300116 detected a clear band at the predicted size (~23 kDa) in WT HAP1 cell lysate, with no corresponding band in Rab35 KO lysate. Additionally, a prominent band of higher molecular weight (>93 kDa) was observed in both test lysates.

In IP, the antibody (2 µg) clearly enriched Rab35 upon precipitation. Rab35 remains detectable in the unbound fraction indicating incomplete 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 weak Rab35-specific staining in HAP1 WT cells, with no background signal in Rab35 KO cells.

We highly encourage other scientists to share their observations.