Messy Staining for Class-II MHC

Pathology
Brigham and Women's Hospital
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

PE Cy7 anti-mouse I-A/I-E Antibody

Catalog Number:

107629

Info: View Product View Product Specs

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Class-II MHC is expressed on all antigen presenting cells including DCs, macrophages and B cells. The expression increases during an immune response. I used this antibody to test MHC-II expression on macrophages. While the expression was positive, it was smeared and messy. On B cells, the expression was very high.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Lymphocytes

Primary Incubation

1:400 in PBS for 30 minutes

Blocking Agent

FcR block

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow Cytometry - PECy7 channel

Results Summary

The expression is positive but non-uniform and smeared. It may be better to titrate or use a different fluorophore.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Very bright staining

The Bad

Smeared and messy

The Bottom Line

Good for staining, but prefer a different fluorophore

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