Human TIM-3 Alexa Fluor® 594-conjugated Antibody

Catalog #: FAB2365T Datasheet
Clone 344823 was used by HLDA to establish CD designation
Catalog # Availability Size / Price Qty
FAB2365T-025
Detection of TIM‑3 in Th1-stimulated Human PBMCs by Flow Cytometry.
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Human TIM-3 Alexa Fluor® 594-conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects human TIM‑3 in direct ELISAs and Western blots. Does not cross-react with recombinant human (rh) TIM‑1, rhTIM‑4, recombinant mouse (rm) TIM‑1, rmTIM‑2, rmTIM‑3, rmTIM‑5, or rmTIM‑6.
Source
Monoclonal Rat IgG2A Clone # 344823
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human TIM‑3
Ser22-Arg200
Accession # Q8TDQ0.2
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 594 (Excitation= 590 nm, Emission= 617 nm)

Applications

Recommended Concentration
Sample
Flow Cytometry
Titration recommended for optimal concentration with starting range of 0.1-1 µg/1 million cells. Samples used for this experiment were Th1-stimulated human peripheral blood mononuclear cells (PBMCs) and PBMC monocytes.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry Detection of TIM-3 antibody in Th1-stimulated Human PBMCs antibody by Flow Cytometry. View Larger

Detection of TIM‑3 in Th1-stimulated Human PBMCs by Flow Cytometry. Unstimulated (A) and Th1-stimulated (B) human peripheral blood mononuclear cells (PBMCs) were stained with Rat Anti-Human TIM-3 Alexa Fluor® 594-conjugated Monoclonal Antibody (Catalog # FAB2365T) and Mouse Anti-Human CD4 APC-conjugated Monoclonal Antibody (FAB3791A). Quadrant markers were set based on control antibody staining (IC006T). View our protocol for Staining Membrane-associated Proteins.

Flow Cytometry View Larger

Detection of TIM‑3 in PBMC monocytes by Flow Cytometry. PBMC monocytes were stained with Rat Anti-Human TIM‑3 Alexa Fluor® 594‑conjugated Monoclonal Antibody (Catalog # FAB2365T, filled histogram) or isotype control antibody (Catalog # IC006T, open histogram). View our protocol for Staining Membrane-associated Proteins.

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

Background: TIM-3

TIM-3 (T cell Immunoglobulin and Mucin domain-3), also known as HAVCR2, is a 60 kDa member of the TIM family of immune regulating molecules. TIMs are type I transmembrane glycoproteins with one Ig-like V-type domain and a Ser/Thr-rich mucin stalk region (1, 2). Mature human TIM-3 consists of a 181 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment, and a 78 aa cytoplasmic tail (3). An alternatively spliced isoform is truncated within the mucin-like stalk. Within the ECD, human TIM-3 shares 58% aa sequence identity with mouse and rat TIM-3. TIM-3 is up‑regulated on several populations of activated myeloid cells (macrophages, monocytes, dendritic cells, microglia, and mast cells) and T cells (Th1, CD8+, NK, and Treg) (3-10). Its binding to Galectin-9 induces a range of immunosuppressive functions which enhance immune tolerance and inhibit anti-tumor immunity (11). TIM-3 ligation attenuates CD8+ and Th1 cell responses (11-13) and promotes the activity of Treg and myeloid derived suppressor cells (8, 11, 13, 14). In addition, dendritic cell-expressed TIM-3 dampens inflammation by enabling the phagocytosis of apoptotic cells and the cross-presentation of apoptotic cell antigens (4). It also binds the alarmin HMGB1, thereby preventing the activation of TLRs in response to released tumor cell DNA (7). TIM-3 interactions with Galectin-9 can alternatively trigger immune stimulatory effects, such as the coactivation of NK cell cytotoxicity (10).

References
  1. Sakuishi, K. et al. (2011) Trends Immunol. 32:345.
  2. Anderson, A.C. (2012) Curr. Opin. Immunol. 24:213.
  3. Monney, L. et al. (2002) Nature 415:536.
  4. Nakayama, M. et al. (2009) Blood 113:3821.
  5. Anderson, A.C. et al. (2007) Science 318:1141.
  6. Wiener, Z. et al. (2007) J. Invest. Dermatol. 127:906.
  7. Chiba, S. et al. (2012) Nat. Immunol. 13:832.
  8. Sanchez-Fueyo, A. et al. (2003) Nat. Immunol. 4:1093.
  9. Ndhlovu, L.C. et al. (2012) Blood 119:3734.
  10. Gleason, M.K. et al. (2012) Blood 119:3064.
  11. Zhu, C. et al. (2005) Nat. Immunol. 6:1245.
  12. Sakhdari, A. et al. (2012) PLoS ONE 7:e40146.
  13. Sabatos, C.A. et al. (2003) Nat. Immunol. 4:1102.
  14. Dardalhon, V. et al. (2010) J. Immunol. 185:1383.
Long Name
T Cell Immunoglobulin Mucin-3
Entrez Gene IDs
84868 (Human); 171285 (Mouse); 363578 (Rat); 479318 (Canine); 102141722 (Cynomolgus Monkey)
Alternate Names
CD366; FLJ14428; HAVCR2; HAVcr-2; hepatitis A virus cellular receptor 2; kidney injury molecule-3; KIM-3; SPTCL; T cell immunoglobulin mucin 3; T cell immunoglobulin mucin-3; T-cell immunoglobulin and mucin domain-containing protein 3; TIM 3; TIM3 T-cell membrane protein 3; TIM3; TIM-3; TIMD3; TIMD-3; TIMD3KIM-3

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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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