Anti-CD8-APC-H7 Ab For The Detection & Characterization Of Human T-Cells By Flow Cytometry

February 24, 2017

Clinical Research
Albert Einstein College of Medicine
Pre-doctoral researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

Mouse anti-human CD8-APC-H7 Ab

Catalog Number:

560179

For our clinical studies, we identify, characterize, and quantify different T-cell sub-populations using a whole blood staining protocol for flow cytometry. Our T-cell panel includes the usual suspects (CD45, CD3, CD4, CD8, etc.), and we stain with this particular CD8 antibody to differentiate between the two major populations of T-cells, CD4 and CD8 T-cells (Plot CD4 vs. CD8 once CD3+CD45+ (total T-cells) have been identified). BD's anti-CD8-APC-H7 antibody provides excellent staining of our study samples.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood (100 ul)

Primary Incubation

Incubate 100 ul whole blood with 5 ul of anti-CD8-APC-H7 Ab for 20 minutes on ice.

Blocking Agent

Incubate whole blood with 10 ul of human serum for 10 minutes on ice prior to primary incubation.

Secondary Incubation

Add RBC lysis buffer, vortex, and incubate (covered/no light) at room temperature for 15 minutes.

Tertiary Incubation

Add DAPI at 1:10 and acquire data immediately.

Detection

Positive staining can be detected using a flow cytometer (we use an LSR-II).

Results Summary

Following data acquisition, analyses can be performed using flow cytometry software such as FlowJo. Make sure to run single color and FMO controls with each experiment to ensure the utmost accuracy of results. Once total T-cells have been identified as a cell population (CD3+CD45+), we plot CD4 vs. CD8 to separate our two main sub-populations prior to further characterization (see image).

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Relatively inexpensive, easy to optimize, simple protocol, reliable and consistent results.

The Bad

Make sure to titrate/optimize antibody with your sample type prior to use on precious samples/materials.

The Bottom Line

Similar to all of our BD flow cytometry antibodies, I highly recommend this product for use in whole blood staining protocols designed to identify T-cells. The read-out is clean, and the staining is consistent across multiple, different sets of experiments.

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