Anti-Sigma 1 Receptor (Western Blot)

Anesthesiology
University of Wisconsin-Madison
Research Associate

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Company:

Santa Cruz Biotechnology

Product Name:

Anti-Sigma 1 Receptor

Catalog Number:

sc-166392

This antibody was used to detected human Sigma 1 Receptor-HA fusion protein expression in overexpressing transfected HEK293 cells. Western blot of HEK293 cells overexpressing the Sigma 1 Receptor-HA fusion protein was performed to compare to the staining pattern using an anti-HA antibody. Bands of the same size proved the anti-Sigma 1 Receptor was capable of the detecting Sigma 1 Receptor protein. This antibody also detected endogenous HEK293 protein. This work was published in "Mavlyutov et al. Sigma-1 receptor expression in the dorsal root ganglion: Reexamination using a highly specific antibody, Neuroscience 331: 148-157."

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293 cell lysate of transfected cells overexpressing human Sigma 1 Receptor-HA fusion protein. 20 ug was reduced, denatured, and loaded onto a SDS-PAGE gel.

Primary Incubation

A dilution of 1:1000 of anti-Sigma 1 Receptor in 2% DGS was incubated over night at 4C.

Blocking Agent

5% milk

Secondary Incubation

Anti-Mouse-HRP (1:5000) in 2% milk incubated for 1 hour at room temperature.

Tertiary Incubation

N/A

Detection

SuperSignal West Femto ECL Substrate (Thermo #34094)

Results Summary

The anti-Sigma 1 Recetor antibody resulted in very strong band at a similar size as the anti-HA staining of the same HEK293 lysate. Additionally anti-Sigma 1 Receptor was able to detect endogenous HEK293 Sigma 1 Receptor protein.

Additional Notes

The top band is the exogenous Sigma 1 Receptor-HA fusion protein and the bottom band is the endogenous Sigma 1 Receptor from HEK293 cells.

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Summary

The Good

The anti-Sigma 1 Receptor was abel to detect exogenous and endogenous Sigma 1 receptor protein.

The Bad

There were some non-specific bands detected.

The Bottom Line

I would cautiously recommend this antibody strictly to be used for western blot analysis however "Mavlyutov et al. Sigma-1 receptor expression in the dorsal root ganglion: Reexamination using a highly specific antibody, Neuroscience 331: 148-157" showed that this antibody produced a non-specific immunoreactive signal in immunohistochemical analysis using Sigma 1 Receptor knockout mice tissue.

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