Immunofluorescence With Anti-MHC II

August 19, 2015

Pathology
Stanford
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Anti-mouse I-A/I-E

Catalog Number:

107601

Info: View Product View Product Specs

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The aim of the research in which this image was procured was to study the role of macrophages in the intestine in both normal and diseased states. Specifically, the goal was to see if cells labeled in the CX3CR1-GFP mouse were different in normal vs diseased mice. This particular product was tested for immunofluorescence because of its robust staining via flow cytometry.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Frozen section from normal, healthy mouse small intestine prefixed in PFA

Primary Incubation

1:100 in 1% BSA in PBS overnight at 4 degrees

Blocking Agent

5% goat serum in 1% BSA in PBS

Secondary Incubation

Goat anti-rat AlexaFluor594 was used at 1:200 in PBS and was incubated on the section at 1 hour at room temperature in the dark

Tertiary Incubation

None

Detection

Zeiss LSM 700 confocal

Results Summary

The antibody worked well. As you can see, most of the MHC II-staining cells in the intestine are in the lamina propria and are additionally marked by the CX3CR1-GFP label (which is expected for these macrophages/mononuclear phagocytes). No staining is detected in the epithelial compartment or by the smooth muscle cells.

Additional Notes

Green here is the CX3CR1-GFP label, red is MHC II, and blue is DAPI.

Related Categories

Image Gallery

Summary

The Good

The clone is great for both flow cytometry and immunofluorescence and stains really well. Also works across multiple mouse backgrounds.

The Bad

Nothing to mention.

The Bottom Line

The product is great to have handy if immunofluorescence or flow cytometry on MHC II+ cells in mice is necessary.

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