FACS Staining of RSV Infected PBMCs

December 11, 2015

Center for Infectious Diseases Control (CIb)
National Institute for Public Health and the Environment (RIVM)
Project Leader

Overall

Quality of Results

Ease-of-Optimization

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Company:

EMD Millipore, Chemicon®

Product Name:

Anti-RSV Antibody, glycoprotein, all type A, B strains, clone 131-2G

Catalog Number:

MAB858-2F

RSV infected PBMCs were detected using this FITC-conjugated monoclonal antibody for extracellular staining.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Fresh human PBMC

Primary Incubation

45 minutes incubation in the dark at ambient temperature

Blocking Agent

PBS supplemented with 0,5% bovine serum albumin

Secondary Incubation

NA

Tertiary Incubation

Fixation by 3.7% Formalin in PBS supplemented with 1% bovine calf serum

Detection

BD FACS Canto II with FACS Diva software

Results Summary

After overnight incubation or up to 24 h post-infection with RSV, infected cells are detected. We found the optimal dilution to be 1:1000.

Additional Notes

None

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Summary

The Good

High dilution factor can be used

The Bad

Separation from negative cells could be better

The Bottom Line

This antibody is more expansive than comparable products, but definitely worth the price in terms of specificity, background, and working concentration.

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