FITC-Conjugated Anti-Rat CD3 (Clone G4.18) Is Not Cross-Reactive With Human CD3FITC-Conjugated Anti-Rat CD3 (Clone G4.18) Is Not Cross-Reactive With Human CD3

January 30, 2017

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

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Company:

BD Pharmingen

Product Name:

FITC Mouse Anti-Rat CD3 clone G4.18

Catalog Number:

554832

Our lab routinely tests antibodies for cross-reactivity to other species for use in immunological assays. This antibody which recognizes rat CD3 exhibits minimal cross-reactivity to human CD3.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human peripheral blood mononuclear cells

Primary Incubation

FITC mouse anti-rat CD3 antibody (BD 554832), incubated with approximately 1x10^6 cells for 1 hour on ice at 1:50 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

As shown in the histogram figure, anti-rat CD3 (clone G4.18) did not exhibit significant cross-reactivity with human lymphocytes (solid black line histogram). It should be noted that some binding was observed to a small minority of the cells (<5%). Cells with a matched isotype control are represented by the grey filled histogram.

DOI or PMID #

N/A

Additional Notes

Gating was performed on the lymphocyte population according to forward vs side scatter for analysis.

Related Categories

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Summary

The Good

Comes directly labelled with FITC. Although not shown here, likely works well against rat tissue.

The Bad

For those working with human tissue, exhibits minimal cross-reactivity (<5%).

The Bottom Line

If you are in urgent need of an antibody against human CD3 for your experiments and you have access to this anti-rat CD3 (clone G4.18) antibody, it will not work as it does not demonstrate cross-reactivity. You are better off using one of the several anti-human CD3 commercially available antibodies.

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