Excellent Antibody Mixture To Detect Human Cytokeratins

May 30, 2018

The Cell and Molecular Biology
The Scripps Research Institute
Assistant Professor

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Company:

MilliporeSigma (Previously Sigma-Aldrich)

Product Name:

Monoclonal Anti-Cytokeratin, pan (mixture)

Catalog Number:

C2562

This reagent (ascites fluid) is a mixture of 6 mouse monoclonal antibodies recognizing human cytokeratins 1, 4, 5, 6, 8, 10, 13, 18, and 19. We successfully used this reagent to identify by IHC human colon carcinoma cells within chick embryo tissues.

Experimental Design and Results Summary

Applications

Immunohistochemistry

Sample

Human colon carcinoma cells within chick embryo chorioallantoic membrane.

Primary Incubation

Tissue was fixed in Zn-formalin and paraffin embedded. Sections were treated with 0.3% hydrogen peroxide and blocked with 2% BSA and 5% goat serum Antigen retrieval was performed with pepsin (Dako). Sections were stained with primary antibodies at a final dilutionof 1:1000, overnight, at 4oC.

Blocking Agent

5% goat serum and 2% BSA in PBS

Secondary Incubation

anti-mosue biotinylated antibody, 1 hr, RT.

Tertiary Incubation

Avidin-HRP.

Detection

DAB chromogenic substrate; counerstaining with hematoxylin.

Results Summary

IHC staining with this anti-keratin antibody reagent produced intense brown staining against light background of the host (chick embryo) tissue.

DOI or PMID #

N/A

Additional Notes

The image depicts the colony of human colon carcinoma cells (stained brown) within the CAM tissue. The final dilution of ascites fluid should be determined for a particular application.

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Summary

The Good

Intense staining that appears highly specific to human carcinoma cells.

The Bad

None detected. Higher background staining could be expected with non-avian host tissues.

The Bottom Line

This anti-human keratin antibody mixture is highly recommended for IHC visualization of pan-cytokeratins in human cells of epithelial origin.

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