FITC-Conjugated Monoclonal Mouse Anti-Human CD3 Antibody (for Flow Cytometry) from BioLegend (Clone UCHT1)

May 23, 2016

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

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Company:

BioLegend

Product Name:

FITC anti-human CD3 Antibody

Catalog Number:

300406

The monoclonal mouse anti-human CD3 antibody, clone UCHT1, is one of the more established monoclonals used for CD3 staining of T cells. We routinely use this antibody to identify the CD3+ T cell population in human PBMC using flow cytometry. This FITC-conjugated version of UCHT1 gave excellent results with bright staining of CD3+ lymphocytes.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC isolated from blood by standard Ficoll density gradient separation

Primary Incubation

FITC anti-human CD3 antibody (BioLegend 300406), incubated with approximately 1x10^6 cells for 1 hour on ice at 1:20 dilution in a total volume of 100ul PBS+2% FCS, 1mM EDTA, 0.1% sodium azide.

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

FACSCalibur (Becton Dickinson)

Results Summary

CD3+ cells were brightly stained and clearly separated from CD3- cells in FACS analysis.

Additional Notes

For flow cytometric analysis, gating was performed on the lymphocyte population. Monocytes were negative (not shown).

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Summary

The Good

Bright staining. Excellent pricing. Available in several test sizes. Well established protocol provided by the manufacturer resulting in quick and simple cell staining. The epitope recognized by UCHT1 is on the surface of cells, therefore fixation and permeabilization of cells is not need which is a plus.

The Bad

None

The Bottom Line

Get this antibody if you plan on staining for CD3 in human PBMC by flow cytometry. Very economical!

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