Superb iCos Ligand Antibody for Identifying Co-Stimulatory Molecules by Flow Cytometry

Department of Immunology and Microbiology
Scripps Research
Graduate Student

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Company:

BioLegend

Product Name:

PE anti-mouse CD275 (B7-H2, B7-RP1, ICOS Ligand) Antibody

Catalog Number:

107405

Clone Number:

HK5.3

The PE iCos Ligand antibody works well for identifying iCos Ligand expression on myeloid immune cells from cell populations isolated from mouse lungs and bronchoalveolar lavage fluid. I found this antibody to provide clear shifts at 1:100-1:200 dilution.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Mouse BALF, Lungs

Primary Incubation

1:100, 30 mins at 4 degrees C in the dark

Blocking Agent

Fc Shield

Secondary Incubation

BD cytofix/cytoperm; 100 ul for 20 mins at 4 degrees C

Tertiary Incubation

None

Detection

5 laser Aurora Flow cytometer

Results Summary

I used this product to identify shifts in iCos ligand expression on dendritic cells, alveolar macrophages and monocytes isolated from the lung 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. I have not seen any spillover issues with this antibody and fluorophore combination.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Specific, bright, and versatile for numerous panels.

The Bad

Dilution of 1:100.

The Bottom Line

I would highly recommend the PE ICos Ligand antibody for identifying shifts in co-stimulatory molecule expression in cells.

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