T cell immunoglobulin mucin-3 (Tim3) is a member of the T cell Immunoglobulin- and Mucin-domain-containing family of type I membrane glycoproteins that regulate autoimmune and allergic disease. Tim3 is selectively expressed on Th1 cells and interacts with galectin-9. It negatively regulates Th1 responses and affects macrophage activation. The 280aa mature human Tim3 contains a V-type Ig-like domain that shows multiple polymorphisms, followed by a mucin-like domain in the 171aa extracellular region, which shares 60% aa identity with the mouse Tim3 ECD. One splice variant is truncated within the mucin domain and presumably is secreted.
Applications
Suitable for use in Flow Cytometry. Other applications not tested.
Recommended Dilution
Flow Cytometry: 10ul labels up to 1x10e6 cells. Validated using whole blood monocytes. Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol
1. Cells may be Fc-blocked with 1ug of human IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction. 2. After blocking, add 10ul of T5469-92F to up to 1x10e6 cells and incubate for 30 minutes at RT 3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (Catalog #F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer. 4. Resuspend the cells in Flow Cytometry Staining Buffer for final flow cytometric analysis. As a control for this analysis, cells in a separate tube should be treated with APC-labeled rat IgG2A antibody.
Storage and Stability
Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: APC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Immunogen
Recombinant protein corresponding to Ser22-Arg200 from the extracellular domain of human Tim3, expressed in NSO cells.
Form
Supplied as a liquid in saline, 0.5% BSA, 0.09% sodium azide. Labeled with Allophycocyanin (APC).
Purity
Purified by Protein G affinity chromatography from hybridoma culture supernatant.
Specificity
Recognizes human Tim3.
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