Myc-Tag Antibody to Detect the Expression of Myc -Tagged Proteins

None
Institute of Protein Biochemistry, Italian National Research Council, Napoli
Postdoctoral fellow

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Company:

Cell Signaling Technology

Product Name:

Myc-Tag Antibody

Catalog Number:

2272

Detection of proteins which have no available antibody are made easy by epitope tags. These tags allow the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunofluorescence. The Myc-tag antibody (#2272) was used successfully to detect myc-tagged RNF215 protein.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa cell lysate overexpressing myc-tagged RNF215 (80ug total protein)

Primary Incubation

Primary antibody (1:1000) in dilution buffer (TBST with 5% BSA) with gentle agitation overnight at 4°C.

Blocking Agent

TBST with 5% w/v nonfat dry milk

Secondary Incubation

Incubated the membrane with the Goat Anti-Rabbit IgG, H & L Chain Specific Peroxidase Conjugate (Cat No: 401315) 1:10000 diluted in 10 ml of blocking buffer with gentle agitation for 1 hr at room temperature.

Tertiary Incubation

None

Detection

Incubated the membrane with appropriate amount of Luminata Crescendo Western HRP substrate (WBLUR0100 Merckmillipore) for 5 min at room temperature. Drained the membrane of excess developing solution (without letting it to dry) wrapped in transparent plastic sheet and exposed the x-ray film for 1 min.

Results Summary

Antibody detects the myc-tagged RNF215 expression in the Hela cells.

Additional Notes

This antibody can be used for immunoflurescence .

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Summary

The Good

Works for both immunofluorescence and western blot

The Bad

None

The Bottom Line

This antibody works better than many antibodies available for the myc-tag.

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