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a-mouse LAG-3 PerCP-Cy5.5
125212
C9B7W
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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.
Flow Cytometry
liver cells infected C57BL/6
1:50 Surface staining, 20 min, 4°C, in 50ul, in the dark
Fc Block
N/A
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.
Cheap, easy to use.
Easy to use antibody that I used in multiple experiments for both analysis and sorting.