Rab1B, Novus Biologicals, NBP3-18251

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Rab1A and Rab1B and followed up by extraction and evaluation of the available data to assess its performance according to pAbmAbs rating criteria.

YCharOS is here conducting a large head-to-head study comparing 12 commercial research antibodies against human Rab1A and Rab1B.

The collection of antibodies was tested using HAP1 WT, RAB1A KO and RAB1B KO cells in the applications WB (WT and KO cell lysates/30 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

In WB (1:500 Ab dilution), NBP3-18251 detects a smeared signal with multiple bands. The most intense band appears above the 57 kDa marker and is observed in all test lysates including lysate from Rab1B KO cells. A weak band can be observed in WT and Rab1A KO cell lysates just below the 24 kDa marker band. The band is absent from Rab1B KO cell lysate, indicating that the antibody specifically detects Rab1B over Rab1A. However, the background signal is high, resulting in a low signal-to-noise ratio and reduced clarity of the specific signal. The antibody (2 µg) does not immunoprecipitate Rab1B from HAP1 WT cell lysates. Rab1B in the starting material, unbound fraction, and precipitate was detected by WB using 17824-1-AP. IF was performed on WT and Rab1B KO cells. An intense signal was observed in WT cells at a 1:100 Ab dilution, with some background signal in KO cells. Additional background originating from the plate was also observed.

We highly encourage other scientists to share their observations.