Anti-mouse CD3 FITC (clone 17A2) is not cross-reactive with human CD3

October 19, 2016

BlueskyReddit
Division of BioMedical Sciences, Faculty of Medicine
Memorial University of Newfoundland
Research Assistant

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Company:

affymetrix eBioscience

Product Name:

Anti-Mouse CD3 FITC (Clone: 17A2)

Catalog Number:

11-0032-80

One of the general aims of our lab is to test for cross-reactivity of commercial available antibodies. This rat anti-mouse CD3 antibody was tested for cross-reactivity to human CD3.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human peripheral blood mononuclear cells

Primary Incubation

FITC-labelled rat anti-mouse CD3 antibody (eBioscience 11-0032-80), incubated with approximately 1x10^6 cells for 1 hour on ice at 1:100 dilution in a total volume of 100ul FACS buffer (PBS+2% FCS, 1mM EDTA, 0.1% sodium azide)

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

FACSCalibur (Becton Dickinson)

Results Summary

This antibody is not cross-reactive to human CD3 as evidenced by a lack of binding to human peripheral blood mononuclear cells as indicated by the black line in the histogram. The grey filled histogram represents the matched isotype control.

Additional Notes

For flow cytometric analysis, gating was performed on the lymphocyte population.

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Summary

The Good

Comes directly labelled with FITC and relatively cheap. Although not shown here, likely works well against mouse tissues.

The Bad

Does not recognize human CD3

The Bottom Line

If you are in an immediate need of an antibody against human CD3 and the only one you have on hand is this product, do not bother as it does not recognize human CD3. It is best to find a validated anti-CD3 antibody for human tissue (which should be no problem given the several that are commercially available).

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