Rab27B, Thermo Fisher Scientific, PA5-54096

Our team performed a comprehensive review of the scientific literature to identify studies using antibodies against human Rab27A and Rab27B, followed by extraction and evaluation of available data according to pAbmAbs rating criteria.

YCharOS conducted a systematic head-to-head comparison of 14 commercially available antibodies against Rab27A and 7 against Rab27B across multiple applications

Antibodies were tested by WB using lysates (40 µg total protein) from U-87 MG WT cells and cells with siRNA-mediated knockdown (KD) of Rab27A or Rab27B. The same cell lines were used for IF testing. In addition, antibodies were evaluated by IP using HAP1 (Rab27A) or U-87 MG (Rab27B) cell lysates (1 mg), following YCharOS standard protocols (see reference).

In WB (1:1000 antibody dilution), PA5-54096 detects a strong band at the expected size (~24 kDa) in WT U-87 MG WT lysate, with no corresponding band in lysate from Rab27B KD cells. A few faint background bands observed.

In IP, the antibody (2 µg) enriches the Rab27B signal compared to the starting material (U-87 MG cell lysate), with a slightly reduced signal observed in unbound fraction, indicating incomplete depletion of the target from the cell lysate. Rab27B in the starting material, unbound fraction, and precipitate was detected by WB using the Rab27B-specific the antibody 17572 from Cell Signaling Technology.

In IF, the antibody (1:200 antibody dilution) resulted in a strong signal from U-87 MG WT with no background signal in Rab27B KD cells.

We highly encourage other scientists to share their observations.