Scavenger Receptor BI (SR-BI) Antibody (for Western Blotting) from Novus Biologicals, Inc.

March 13, 2013

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Medical Biochemistry
University of Cape Town, South Africa
Research Officer
Western Blot assessing expression of SR-B1 in different macrophage populations derived from wild type C57/BL6 and SR-B1-/- mice (from: Schäfer et al., PLoS one 4 (12): e8448).

Company:

Novus Biologicals, Inc.

Product Name:

Scavenger Receptor BI (SR-BI) antibody

Catalog Number:

NB 400-104

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 400-104) gave the best results in Western blotting.

Experimental Design and Results Summary

Applications

Western blotting

Sample

Whole cell extracts of cultured mouse cells (NIH3T3 fibroblasts and RAW macrophages) and different primary mouse macrophage populations (alveolar, bone-marrow-derived, resident peritoneal and thioglycollate-elicited peritoneal); homogenized tissue (rat liver)

Primary Incubation

SR-BI antibody ( Novus Biologicals, cat.no. NB 400-104); overnight incubation at 4°C in blocking reagent (1:1000 dilution)

Blocking Agent

5% milk powder in PBS-Tween

Secondary Incubation

Anti-rabbit IgG HRP conjugate (Jackson Immuno Research Laboratories, cat. no. 211-035-109), 1h incubation at room temperature; dilution 1:10.000 in blocking reagent

Tertiary Incubation

N/A

Detection

ECL chemiluminescence detection reagents (GE Life Sciences); detection with the VisionWorks LS Biospectrum™500 Imaging System (UVP).

Results Summary

This antibody worked excellently in Western blotting with highly specific detection of the 82kDa SR-BI protein without background staining. It detects both mouse and rat SR-BI.

Additional Notes

Of all the SR-BI antibodies tested (I also tested Novus Biologicals NB 400-113 and NB 400-134), only this one (Novus Biologicals NB 400-104) gave high quality Western blot results. However, this antibody could not be successfully used for other applications such as FACS, immunostaining or functional receptor blocking experiments. It is also worthwhile to note that after primary antibody incubation, the antibody in the blocking agent could be stored at -20°C and reused several times.

Image Gallery

Western Blot assessing expression of SR-B1 in different macrophage populations derived from wild type C57/BL6 and SR-B1-/- mice (from: Schäfer et al., PLoS one 4 (12): e8448).

Summary

The Good

High quality Western blots, highly specific, re-usable.

The Bad

Only useful in Western blotting.

The Bottom Line

This is a very good antibody to detect SR-BI in cell lysates of different origins. However, for applications other than Western blotting, a whole range of different SR-BI antibodies is needed, each being specific for a particular application.

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