Tubulin alpha-4A chain, Abcam, ab177479

Our team performed a comprehensive review of the scientific literature to identify studies using antibodies against human Tubulin alpha-4A chain, 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 Tubulin alpha-4A chain across multiple applications.

Antibodies were tested by WB using lysates (50 µg protein) from HAP1 and HeLa WT and Tubulin alpha-4A chain-deficient (TUBA4A KO) HAP1 and HeLa cells. These HAP1 WT and KO cells were also used for IF. In addition, antibodies were evaluated by IP using HAP1 cell lysate (0.5 mg protein). All experiments were performed according to YCharOS standard protocols (see reference).

In WB (1:1000 antibody dilution), ab177479 detected a single band of the expected size (~ 50 kDa) in HAP1 and HeLa WT lysates, with only a faint corresponding band in the TUBA4A KO cell lines.

In IP, the antibody (2 µg) clearly enriched Tubulin alpha-4A chain from HAP1 lysate, with residual target protein detected in the unbound fraction, indicating incomplete precipitation. Tubulin alpha-4A chain protein levels in the starting material, unbound fraction, and precipitate were assessed by WB using the Tubulin alpha-4A chain-specific antibody MA5-32738 from Thermo Fisher Scientific (1:5000).

In IF, the antibody (1:150 dilution) failed to produce a Tubulin alpha-4A chain-specific signal in HAP1 WT cells.

We highly encourage other scientists to share their observations.