Rab27B, Synaptic Systems, 168 103

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), 168 103 detects a single band at the expected size (~24 kDa) in WT U-87 MG WT lysate, with no corresponding band in lysate from Rab27B KD cells.

In IP, the antibody (2 µg) enriches the Rab27B signal compared to the starting material (U-87 MG cell lysate), with a 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 (1:1000 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.

This antibody works well for immunostaining Rab27B in primary hippocampal neurons from mice. 7 DIV neurons were fixed in 4% PFA and immunostained for Rab27B using 1:500 dilution in blocking buffer. Little background was observed.