Rab7A, Abcam, ab137029

Our team has carried out deep research in the published scientific literature to identify papers and reports using antibodies directed against human Rab7A 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 14 commercial research antibodies against human Rab7A encoded by RAB7A.

The collection of antibodies was tested using HeLa WT and RAB7A KO cells in the applications WB (WT and KO cell lysates/40 µg protein), IP (WT cell lysate) and IF (WT and KO cells) according to YCharOS standard protocols (see reference).

In WB (1:1000 ab dilution) ab137029 detected a band of appr. 24 kDa in WT cell lysate with no corresponding band observed in KO cell lysate. A band was detected following IP (2 µg Ab used), however this band had same intensity as band in input extract (4% of starting material) and immunodepleted extract (4% of total unbound fraction), showing that the antibody does not deplete the antigen from the cell lysate. In IF (1:100 ab dilution), an intense staining was observed in WT cells compared to KO cells showing no background.

We highly encourage other scientists to share their observations.

This antibody works well for immunofluorescence staining in mouse (b.End3) and in human (hCMEC/D3) brain endothelial cell line. Dilution 1:200. Fixation was done with methanol cells were permeabilized with 0.25% Triton X-100.

Producer: Abcam
Catalog No: ab137029
Used dilution: 1:1000 (WB)
Secondary Ab: Anti-Rabbit HRP, Cell signaling 7074 (1:2000)
Cell line: primary porcine brain endothelial cells, bEND3
Host organism: Rabbit
Character: primary, monoclonal

Antibody tested with primary porcine and bEND3 cell lysis.
In case of the primary porcine cell lysis the result was unclear, and multiple bands were detected.
The bEND3 cell lysis produced a single strong band at the appropriate size.

Producer: Abcam
Catalog No: ab137029
Used dilution: 1:100 (IF)
Cell line: primary porcine brain endothelial cells, primary bovine brain endothelial cells
Host organism: Rabbit
Character: primary, polyclonal

Antibody tested in primary porcine and bovine brain endothelial cells. In both cases working very good giving strong and clear dot-like signal. For IF as a secondary antibody Invitrogen goat anti-mouse Alexa Fluor 568 (1:350 or 1:200) was used. Cells fixed with 4% PFA, permeabilized with 0.1% Triton-X, blocked with 2% BSA.