Mouse TIM-3 Antibody

Catalog # Availability Size / Price Qty
AF1529
AF1529-SP
Detection of Mouse TIM‑3 by Western Blot.
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Product Details
Citations (3)
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Mouse TIM-3 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse TIM-3 in direct ELISAs and  Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human TIM‑3 is observed and less than 5% cross-reactivity with recombinant mouse (rm) TIM‑1, rmTIM‑2, rmTIM‑4, rmTIM‑5, rmTIM‑6, and rmTIM‑7 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse TIM‑3
Leu22-Arg191
Accession # AAL65156
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Flow Cytometry
2.5 µg/106 cells
HT‑2 mouse T cell line
Immunohistochemistry
3-15 µg/mL
Immersion fixed paraffin-embedded sections of mouse spleen
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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

Western Blot Detection of Mouse TIM-3 antibody by Western Blot. View Larger

Detection of Mouse TIM‑3 by Western Blot. Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Mouse TIM-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1529) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for TIM-3 at approximately 45-70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry View Larger

TIM‑3 in Mouse Spleen. TIM‑3 was detected in immersion fixed paraffin-embedded sections of mouse spleen using Goat Anti-Mouse TIM‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1529) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: TIM-3

TIM-3 (T cell immunoglobulin and mucin domain-3) 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 (1-3). There are three TIM genes in human and eight in mouse. Mature mouse TIM-3 consists of a 174 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment (TM), and a 67 aa cytoplasmic tail (4). Two alternately spliced isoforms have been reported in mouse which lack either the TM or both the TM and mucin regions (5, 6). Within the ECD, mouse TIM-3 shares 58% and 74% aa sequence identity with human and rat TIM-3, respectively. TIM-3 is specifically expressed on Th1 cells whereas TIM-1 and TIM-2 are expressed on Th2 cells. In chronic inflammation, autoimmune disorders, and some cancers, TIM-3 is upregulated on several other hematopoietic cell types and on hippocampal neurons (9-12). The glycosylated Ig domain of TIM-3 binds cell-associated galectin-9 which induces TIM-3 Tyr phosphorylation and proapoptotic signaling (10, 13). TIM-3 functions as a negative regulator of Th1 cell activity. Its blockade results in increased IFN-gamma production, Th1 cell proliferation, and cytotoxicity (5, 7, 12, 14). TIM-3 may play a role in regulatory T cell development (7), inflammation (15), and immune tolerance (5, 13, 14). Soluble mouse TIM-3 has been shown to inhibit anti-tumor effector T cell responses and to enhance autoimmune reactions (6, 7).

References
  1. Anderson, A.C. and D.E. Anderson (2006) Curr. Opin. Immunol. 18:665.
  2. Mariat, C. et al. (2005) Phil. Trans. R. Soc. B 360:1681.
  3. Meyers, J.H. et al. (2005) Trends Mol. Med. 11:362.
  4. Monney, L. et al. (2002) Nature 415:536.
  5. Sabatos, C.A. et al. (2003) Nat. Immunol. 4:1102.
  6. Geng, H. et al. (2006) J. Immunol. 176:1411.
  7. Sanchez-Fueyo, A. et al. (2003) Nat. Immunol. 4:1093.
  8. Khademi, M. et al. (2004) J. Immunol. 172:7169.
  9. Wiener, Z. et al. (2007) J. Invest. Dermatol. 127:906.
  10. van de Weyer, P.S. et al. (2006) Biochem. Biophys. Res. Commun. 351:571.
  11. Gielen, A.W. et al. (2005) J. Neuroimmunol. 164:93.
  12. Oikawa, T. et al. (2006) J. Immunol. 177:4281.
  13. Zhu, C. et al. (2005) Nat. Immunol. 6:1245.
  14. Koguchi, K. et al. (2006) J. Exp. Med. 203:1413.
  15. Frisancho-Kiss, S. et al. (2006) J. Immunol. 176:6411.
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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Citations for Mouse TIM-3 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Transcriptomic Hallmarks of Tumor Plasticity and Stromal Interactions in Brain Metastasis
    Authors: E Wingrove, ZZ Liu, KD Patel, A Arnal-Esta, WL Cai, MA Melnick, K Politi, C Monteiro, L Zhu, M Valiente, HM Kluger, VL Chiang, DX Nguyen
    Cell Rep, 2019-04-23;27(4):1277-1292.e7.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  2. The HIF-1/glial TIM-3 axis controls inflammation-associated brain damage under hypoxia.
    Authors: Koh H, Chang C, Jeon S, Yoon H, Ahn Y, Kim H, Kim I, Jeon S, Johnson R, Park E
    Nat Commun, 2015-03-20;6(0):6340.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  3. TIM-1 and TIM-3 enhancement of Th2 cytokine production by mast cells.
    Authors: Nakae S, Iikura M, Suto H, Akiba H, Umetsu DT, DeKruyff RH, Saito H, Galli SJ
    Blood, 2007-07-09;110(7):2565-8.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot

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