CD4 Expression On The PE-Cy7 Channel

Cancer Immunology & AIDS
Dana Farber Cancer Institute
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

eBioscience

Product Name:

PE-Cy7 anti-mouse CD4

Catalog Number:

25-0042-82

This antibody was used as a compensation control for the PE-Cy7 channel. Although it worked it just fine for my intended purpose, there are a few issues I noticed with the staining. First, even at 1:500 dilution the fluorescence was very bright (>3 log shift) which is perhaps an attribute of the PECy7 dye. Second, the negative population showed some background staining which is an issue for experiments that look at changes in MFI values.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse splenocytes

Primary Incubation

25 minutes on ice

Blocking Agent

CD16/32 antibody for blocking FC receptors

Secondary Incubation

None

Tertiary Incubation

None

Detection

Flow cytometry on PE-Cy7 channel

Results Summary

The CD4 positive cells were stained well and showed relevant expression. The negative fraction of cells have some background staining.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Very bright fluorescence

The Bad

Background staining on negative cells

The Bottom Line

A good product for measuring CD4+ cells, but not for measuring MFI changes.

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