Immunodetection of ENO1 Protein in Mouse Cell Line

Physiology
University of Tennessee HSC
Postdoctoral Researcher

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Company:

Abcam

Product Name:

Anti-ENO1 antibody [EPR10863(B)]

Catalog Number:

ab155102

Clone Number:

N/A

To measure ENO1 levels in a mouse cell line treated with a growth factor, mouse cells at the log phase were treated with the growth, and cells were lysed in RIPA buffer. Proteins were extracted, estimated, and resolved in 10% SDS-PAGE. The resolved proteins were then western transferred in a nitrocellulose membrane and probed with ENO1 antibody. The selection of the ENO1 antibody from Abcam was based on our previous experience with the same antibody.

Experimental Design and Results Summary

Applications

Western blot

Sample

Mouse cell line whole-cell protein lysate

Primary Incubation

Dilution (1:1000) in 5% skimmed milk prepared in 1x TBST, overnight incubation at 4 degrees celsius.

Blocking Agent

5% skimmed milk made in 1x TBST.

Secondary Incubation

Goat anti-Rabbit IgG HRP Ab, dilution (1:4000) in 5% skimmed milk made in 1x TBST, 1hr at RT.

Tertiary Incubation

None

Detection

Enhanced chemiluminescent detection in X-Ray films.

Results Summary

A bright and specific band of ENO1 was detected by the Abcam antibody. However, a few more non-specific proteins band were observed. No effect of treatment on study state levels of ENO1 was also found.

DOI or PMID #

None

Additional Notes

None

Related Categories

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Summary

The Good

Detected particular band of ENO1.

The Bad

Other nonspecific bands were detected.

The Bottom Line

Great antibody.

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