Immunodetection of IL-1ß Expression Levels in Mouse Cells

Physiology
University of Tennessee HSC
Postdoctoral Researcher

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Company:

Cell Signaling Technology

Product Name:

IL-1β (D3U3E) Rabbit mAb

Catalog Number:

12703

Mouse macrophage cells were knockdown for specific protein using its specific siRNA, growth arrested and treated with growth factor for specific time point. Cells were lysed in RIPA buffer, protein extract were made, and using western blot tested for expression levels of IL-1β using its specific antibodies. Cell Signaling Technology antibody was selected because of picture provided in antibody flyer.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Mouse macrophage cells extract

Primary Incubation

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

Blocking Agent

5% skimmed milk prepared in TBST

Secondary Incubation

Dilution (1:4000) in 5% skimmed milk prepared in TBST, 1hr at RT

Tertiary Incubation

None

Detection

Enhanced chemiluminiscent detection in X-Ray film

Results Summary

Poor detection of IL-1ß was observed with antibody. However, there were less non-specific bands so; it can be graded as average antibody for IL-1ß in mouse macrophage cells.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Gives IL-1ß band

The Bad

None

The Bottom Line

Average antibody

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