Works in FACS, but Needs High Concentration

Neurosurgery
BWH/HMS
postdoc

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Horizon

Product Name:

V500 Mouse Anti-Human CD8

Catalog Number:

561617

This antibody was used on human PBMC cells stimulated in vitro with OKT3 for 2 and 3 days. For the optimization I used multiple concentrations (from pure to 1:30).

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC

Primary Incubation

30 min

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

FACSCalibur (Becton Dickinson)

Results Summary

5ul of pure antibody are enough to have a "not bad" positive peak. I used this in combination with 5 other colors and I had problems to compensate it.

Additional Notes

None

Related Categories

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Summary

The Good

Poor background

The Bad

Needs high concentration

The Bottom Line

This antibody is not working excellently in flow cytometry, even if PBMC is stimulate with OKT3. Has to be used pure, no dilution.

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