Great antibody for detecting human CD8 expression

January 29, 2015

Cancer Immunology & AIDS
Dana Farber Cancer Institute
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

FITC Anti-human CD8a

Catalog Number:

301006

I have used this antibody for analyzing the fraction of CD8+ve T cells in a human PBMC derived T cell pool. The human T cells were purified from PBMCs using a magnetic bead based separation method, followed by staining with anti-human CD8a mAb. The cells were analyzed using flow cytometry. The staining looks pretty neat with two separate populations of CD8+ve and CD8-ve cells. The background staining in the negative population is minimal. The frequencies are physiologically relevant and thus, it's a recommended antibody.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human T cells purified from PBMCs

Primary Incubation

1:100 dilution for 30 minutes on ice

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometry on FITC channel

Results Summary

The results in the figure show a clear positive and negative population for expression of surface CD8. These frequencies are physiologically relevant and the background is minimal.

Additional Notes

None

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Image Gallery

Summary

The Good

Great specificity

The Bad

The company reccomends 5ul/test. However, I have titrated and found 1:100 dilution to work just fine.

The Bottom Line

It's one of the most inexpensive but highly specific antibodies available for CD8. The company is now making diluted versions of these antibodies and recommends using 5ul/test, which is too much. One should titrate the concentration and perhaps use a lower concentration.

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