Immunoprecipitaiton (IP) Using ITCH Antibody

March 30, 2015

Department of Cancer Genetics and Epigenetics
City of Hope Cancer Research Hospital
Assistant Research Professor

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Quality of Results

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Company:

BD Biosciences

Product Name:

ITCH antibody

Catalog Number:

611199

Browse Similar Products: ITCH Antibodies

Immunoprecipitation of ITCH from KB cell extracts using ITCH (BD Biosciences: Cat# 611199) mouse mAb (lane 1) or mouse IgG isotype Control (lane 2) was performed. Western blot analysis was performed using ITCH (BD Biosciences: Cat# 611199) mouse mAb. The resulting ITCH protein band from IP is very specific.

Experimental Design and Results Summary

Applications

Immunoprecipitation first, then followed by Western blot

Sample

Protein lysates from KB cancer cells

Primary Incubation

5 ug/ml, overnight, 4 degrees

Blocking Agent

Odyssey blocking buffer

Secondary Incubation

1:2000, room temp, 1 hr

Tertiary Incubation

NA

Detection

Western blot

Results Summary

The ITCH protein co-immunoprecipitated by this antibody is very clean and specific.

Additional Notes

None

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Summary

The Good

The background of the resulting IP data is very clean

The Bad

There are some bands lower than 103-110 kDa (the size of ITCH) detected by this antibody in the Western blot. Not sure if they are non-specfic bands or the degraded ITCH proteins

The Bottom Line

This antibody is very good in IP application.

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