Good Anti-Mouse Tim-3 Antibody

Charite Strasse 1
Deutsches Rheuma-Forschungszentrum
Research Scientist

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Company:

BD Biosciences

Product Name:

BD Horizon™ PE-CF594 Mouse Anti-Mouse CD366 (TIM-3)

Catalog Number:

566998

Clone Number:

5D12/TIM-3

In different healthy and pathological contexts, CD4 T cells differentiate into a range of specialized interleukin-secreting subsets. In mouse models of aging and autoimmune diseases, we study the plasticity and effector functions of CD4 T helper cells. We analyzed the expression of the inhibitory molecule Tim-3 on CD4 T cells with an anti-TIM-3 PE-CF594 antibody in old C57Bl/6J mice.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Splenocytes of wild-type 24 months-old C57Bl/6J mouse, incubated in complete culture medium at 37 °C with (Restimulated) or without (unstimulated) PMA/ionomycin for 4h and brefeldin A for the 3 last hours.

Primary Incubation

Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.

Blocking Agent

2.4G2

Secondary Incubation

Surface staining with anti-CD3 BUV805, anti-CD8 FITC, anti-CD4 BUV495, anti-Tim3 PECF594 1/50, etc., in Cell Staining Buffer (Biolegend) incubated 30 min at 4°C.

Tertiary Incubation

Cells were fixed and permeabilized with a commercial kit to counterstain for cytokines.

Detection

Yellow-green laser on Symphony cytometer

Results Summary

Tim-3 was clearly detected on CD3e+CD4+ T cells. The staining was slightly lower on stimulated cells but still well separated.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Relatively bright staining compared with other conjugates.

The Bad

None

The Bottom Line

Very good option, easy to use.

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