Detect Antigen Presentation Of OVA On MHC-1 Using Flow Cytometry

Cancer Immunology & AIDS
Dana Farber Cancer Institute
Research Fellow

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Company:

Biolegend

Product Name:

PE anti-mouse H-2Kb bound to SIINFEKL

Catalog Number:

141604

This antibody specifically reacts with the ovalbumin-derived peptide SIINFEKL bound to H-2Kb of MHC class I (which is present in C57BL/6 mice), but not with unbound H-2Kb or H-2Kb bound with an irrelevant peptide. I used this antibody to stain a transgenic cell line that expresses mouse Kb. The assay worked perfectly and specific expression of SIINFEKL loaded cells was detected using this antibody.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

K562 transgenic cells

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 channel

Results Summary

Specific staining was observed only when cells were pulsed with 1uM SIINFEKL peptide. Non-pulsed cells or irrelevant cells pulsed with SIINFEKL did not show any expression.

Additional Notes

None

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Summary

The Good

Very good staining

The Bad

A bit expensive

The Bottom Line

Although a bit expensive, this antibody facilitates a really good assay to detect antigen presentation.

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