Clearly Labels CD5+ B Cells From Mouse Spleen

December 16, 2016

Immunology
Duke University
Predoctoral

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

PE-Cy5 Rat Anti-Mouse CD5

Catalog Number:

553024

The goal of this study was to evaluate mouse splenic B cell activation molecule expression after short-term in vitro stimulation with different mitogenic stimuli.This product was selected for its reported brightness relative to other CD5 mAbs.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse Splenocytes

Primary Incubation

Isolate splenocytes. Wash cells once in FACS buffer (PBS plus 1% BSA or serum). Block Fc receptors and stain for viability in PBS for 20 minutes on ice.

Blocking Agent

Fc block

Secondary Incubation

Wash cells once in FACS buffer. Stain with isotype control, CD19, CD5, and/or CD1d antibodies (1:500 dilution, 100 uL per million cells) for 20 minutes on ice.

Tertiary Incubation

N/A

Detection

Flow Cytometer

Results Summary

Unstimulated B cells (left panel, gray histogram in right panel) and B cells stimulated with LPS for 5 hours (middle panel, black histogram in right panel) show shifts in CD5 expression versus isotype control (empty histogram in right panel), consistent with what has previously been observed.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Easy to use and shows a shift in fluorescence when compared to isotype.

The Bad

Somewhat dimmer than other CD5-reactive antibodies.

The Bottom Line

It clearly shows a shift in CD5 expression upon cell activation, though if some cells are expressing low levels of CD5, the experiment will require a good isotype control for comparison.

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