Scavenger Receptor BI Antibody (for Flow Cytometry) from Novus Biologicals, Inc.

March 19, 2013

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Medical Biochemistry
University of Cape Town, South Africa
Research Officer
FACS analysis of R6F cells stably transduced with pFB (vector control), SR-B1 or SIGNR1, and stained with anti-SR-B1 or anti-SIGNRI, as indicated (from: Schäfer et al., PLoS one 4 (12): e8448).

Company:

Novus Biologicals, Inc.

Product Name:

Scavenger Receptor BI (SR-BI) antibody

Catalog Number:

NB400-113

Image

This antibody was used in a study that investigated the role of Scavenger Receptor BI (SR-BI) for infection with Mycobacterium tuberculosis. It has been published in PLoS One in 2009 (Schäfer et al., The Role of Scavenger Receptor B1 in Infection with Mycobacterium tuberculosis in a Murine Model. PLoS one 4 (12): e8448). Several SR-BI antibodies were tested in this study for various applications; the antibody reviewed here (NB 400-113) gave best results in FACS staining of SR-BI on the surface of live cells stably transfected with SR-BI.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Live R6F cells (Rat 6 fibroblast, SR-BI negative cell line) stably transfected with murine SR-BI

Primary Incubation

SR-BI antibody (Novus Biologicals, cat.no. NB 400-113), incubated 1h at 4°C in blocking reagent (1:300 dilution)

Blocking Agent

5% heat-inactivated rabbit serum; 0.5% BSA; 2mM NaN3; 5mM EDTA in PBS

Secondary Incubation

R-Phycoerythrin-conjugated anti-rabbit IgG (Jackson Immuno Research Laboratories, cat. no. 711-116-152), incubated 1h incubation at 4°C (1:200 dilution)

Tertiary Incubation

N/A

Detection

FACSCalibur (Becton Dickinson), software CellQuest version 4

Results Summary

This antibody worked excellently in FACS analysis of cells overexpressing SR-BI. Only minimal background staining compared to isotype control could be detected.

Additional Notes

Of all the SR-BI antibodies tested (Novus Biologicals NB 400-104 and NB 400-134) only this one (Novus Biologicals NB 400-113) reviewed here gave satisfying results in FACS analysis. However, this antibody could not be successfully used for other applications such as Western blotting or functional receptor blocking experiments.

Image Gallery

FACS analysis of R6F cells stably transduced with pFB (vector control), SR-B1 or SIGNR1, and stained with anti-SR-B1 or anti-SIGNRI, as indicated (from: Schäfer et al., PLoS one 4 (12): e8448).

Summary

The Good

Highly specific in FACS analysis, low background.

The Bad

Only useful for FACS.

The Bottom Line

This is a very good antibody to detect SR-BI on the surface of cells that overexpress SR-BI. The minimal background staining is tolerable when comparing to overexpressing cells. However, this antibody might not be suitable to detect slight differences in SR-B1 surface expression between different cell populations.

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