Relatively Dim but Versatile MHC Class I Antibody for Flow Cytometric Analysis

Department of Immunology and Microbiology
Scripps Research
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Alexa Fluor® 700 anti-mouse H-2Kb Antibody

Catalog Number:

116522

Clone Number:

AF6-88.5

The Alexaflour 700 H-2kb antibody is effective in identifying MHC class I-expressing cell populations within immune cells isolated from mouse lungs and bronchoalveolar lavage fluid. It can be utilized for myeloid cells, including dendritic cells, monocytes, macrophages, and neutrophils.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Mouse Lungs

Primary Incubation

Extracellular antibodies at 4C; 30 mins

Blocking Agent

Fc Shield

Secondary Incubation

BD cytofix/cytoperm

Tertiary Incubation

None

Detection

5-laser Cytek Aurora

Results Summary

I used this product to identify MHC-1 expressing cells amongst monocytes, dendritic cells, macrophages, and other myeloid cells isolated from the lung / BALF of unimmunized versus intranasally immunized mice. Cells were isolated from BALF using 1 ml of 1% BSA in PBS and from lungs through collagenase digestion; RBC lysis was performed using ACK buffer. Cells were then stained with viability dye followed by extra-cellular antibodies such as CD45, CD90.2, CD19, Siglec-F, CD11b, CD11c, Ly6C, Gr1, MHCII, MHCI.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Versatile fluorophore for various panels

The Bad

None so far.

The Bottom Line

I would recommend the Af700 H-2kb antibody for detection of MHC-I expression in cell populations isolated from the lung, BALF, lymph nodes or spleen.

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