CD3-AF700 Works Great To Detect T Cells In Human PBMC

Medicine
Vanderbilt University Medical Center
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

Alexa Fluor® 700 Mouse Anti-Human CD3

Catalog Number:

561027

The general aim of our research is to characterize the phenotype and function of immune cells in various diseases. We are building fluorescence and mass cytometry panels to study human PBMC in the context of autoimmune and idiopathic pulmonary diseases. The main reason I selected CD3-AF700 was because it is the best CD3 clone and fit well in my 9-parameter panel.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Cryopreserved human PBMC

Primary Incubation

0.5uL (1:400) CD3-AF700 in 200uL FACS buffer with 2x10^6 cells; 15 minutes; room temperature

Blocking Agent

None

Secondary Incubation

None

Tertiary Incubation

None

Detection

BD LSRII 5-laser Flow Cytometer

Results Summary

A single color titration of CD3-AF700 was performed and 0.5uL (1:400) yielded the highest stain index. We routinely use CD3-AF700 in our 9-parameter PBMC panel to identify T cells in patient samples with various diseases. Cyropreserved human PBMCs are thawed, washed twice in PBS, and stained with a cocktail of surface antibodies, including CD3-AF700. ~2 million cells in 200uL FACS buffer are incubated with antibodies for 15 minutes at room temperature.As evidenced in the attached figure, the staining looks great. It detects the expected frequency of T cells in human PBMCs.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Good staining pattern

The Bad

None

The Bottom Line

Antibody works great!

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