Great LAG-3 Antibody for the Detection of Tr1 Cells

Protozoa Immunology
Bernhard Nocht Institut
Research Scientist

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Company:

BioLegend

Product Name:

a-mouse LAG-3 PerCP-Cy5.5

Catalog Number:

125212

Clone Number:

C9B7W

Info:Supplier Page View Product Specs

Browse Similar Products: LAG3 Antibodies

I used this antibody to characterize and sort Tr1 cells (identified as CD49b+ LAG-3+ CD4 T cells) in mice infected with Plasmodium berghei ANKA (PbA), a murine model for malaria.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

liver cells infected C57BL/6

Primary Incubation

1:50 Surface staining, 20 min, 4°C, in 50ul, in the dark

Blocking Agent

Fc Block

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Flow Cytometry

Results Summary

This antibody was used in combination with CD49b to identify Tr1 cells and other regulatory cells in mice, characterized by their LAG-3 expression. Whilst PerCP-Cy5.5 is not my favourite fluorochrome, being difficult to compensate PerCP, PE-Dazzle and some other fluorochromes, I was always able to detect a good signal on my cells when employing good panel design. Furthermore, our sorter (Aria IIIu) is set up to detect 7 fluorochromes, among them PerCP-Cy5.5, and I was happy to use this antibody / fluorochrome as it worked well. This antibody is also fairly cheap, enabling me to use it for sorting cells from multiple pooled spleens, requiring a lot of antibody, at a fairly low cost. In normal ex vivo staining/ analysis, I used it at a dilution of 1:50 in 50ul for 1.5E6 cells. In the following, you will find my staining protocol in short. Incubation steps were performed at 4°C in the dark. 1.Count and adjust splenocytes to 1.5E6 cells, plate in a 96-well plate. 2.If required, stain cells with viability dye. Wash. 3. Stain with extracellular master mix for 20 min. Wash 2x. 4. Resuspend in FACS buffer and acquire as soon as possible. Here, data was acquired on a BD LSR II or Aria IIIu.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Cheap, easy to use.

The Bad

N/A

The Bottom Line

Easy to use antibody that I used in multiple experiments for both analysis and sorting.

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