Human TIM-3 Antibody Summary
Ser22-Arg200
Accession # Q8TDQ0
Applications
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
TIM-3 in Human Tonsil Using Dual RNAscope®ISH and IHC. TIM-3 mRNA was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope®Probe (Catalog # 560681) and stained using ACD RNAscope®2.5 HD Detection Reagents-Red (top image, Catalog # 32260). Adjacent tissue section was processed for immunohistochemistry using R&D Systems Goat Anti-Human TIM-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2365) at 3 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC004) and DAB chromogen (lower image, yellow-brown). Tissues were counterstained with hematoxylin (blue).
TIM‑3 in Human Tonsil. TIM-3 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human TIM-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2365) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes and extracellular space. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Western Blot Shows Human TIM‑3 Specificity by Using Knockout Cell Line. Western blot shows lysates of HDLM‑2 human Hodgkin’s lymphoma cell line and human TIM-3 knockout HDLM‑2 human Hodgkin’s lymphoma cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TIM‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2365) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for TIM‑3 at approximately 50 kDa (as indicated) in the parental HDLM‑2 human Hodgkin’s lymphoma cell line, but is not detectable in knockout HDLM‑2 human Hodgkin’s lymphoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Detection of Human TIM-3 by Immunohistochemistry Immunohistochemical staining of peritoneum tissue for CD4, CD8, CD56, OPN, CD68, Gal-9, and TIM-3. HE, hematoxylin and eosin. Magnification, 20×. Arrows indicate loosen tissue at the broken boundary of the granuloma. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/1422-0067/18/7/1382), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TIM-3 by Simple Western TIM-3 Association with Intracellular Kinases in CD8+/MART-1+ T cells.TIM-3 co-immunoprecipitation analysis of unactivated and 15 min. stimulation with anti-CD3/CD28 beads (activated). Equivalent amounts of protein (~2mg) were co-immunoprecipitated with pAb anti-TIM-3 antibody and western blot was performed using capillary electrophoresis. Cleared lysate served as a loading control for individual antibody reactivity. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0140694), licensed under a CC-BY license. Not internally tested by R&D Systems.
Reconstitution Calculator
Preparation and Storage
- 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 human TIM-3 consists of a 181 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment, and a 78 aa cytoplasmic tail (4). An alternately spliced isoform is truncated following a short substitution after the Ig-like domain. Within the ECD, human TIM-3 shares 58% aa sequence identity with mouse and rat TIM-3. TIM-3 is expressed on the surface of effector T cells (CD4+ Th1 and CD8+ Tc1) but not on helper T cells (CD4+ Th2 and CD8+ Tc2) (4, 5). In chronic inflammation, autoimmune disorders, and some cancers, TIM-3 is upregulated on several other hematopoietic cell types. It also occurs on hippocampal neurons (7-10). The Ig domain of TIM-3 interacts with a ligand on resting but not activated Th1 and Th2 cells (5, 11). The glycosylated Ig domain of TIM-3 binds cell-associated galectin-9. This induces TIM-3 Tyr phosphorylation and pro-apoptotic signaling (8, 12). 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, 10, 11, 13), regulatory T cell development (5), and increases in macrophage and neutrophil infiltration into sites of inflammation (14). Soluble mouse TIM-3 constructs which lack the cytoplasmic domain have been shown to inhibit anti-tumor effector T cell responses and to enhance autoimmune reactions (5, 15).
- Anderson, A.C. and D.E. Anderson (2006) Curr. Opin. Immunol. 18:665.
- Mariat, C. et al. (2005) Phil. Trans. R. Soc. B. 360:1681.
- Meyers, J.H. et al. (2005) Trends Mol. Med. 11:362.
- Monney, L. et al. (2002) Nature 415:536.
- Sanchez-Fueyo, A. et al. (2003) Nat. Immunol. 4:1093.
- Khademi, M. et al. (2004) J. Immunol. 172:7169.
- Wiener, Z. et al. (2007) J. Invest. Dermatol. 127:906.
- van de Weyer, P.S. et al. (2006) Biochem. Biophys. Res. Commun. 351:571.
- Gielen, A.W. et al. (2005) J. Neuroimmunol. 164:93.
- Oikawa, T. et al. (2006) J. Immunol. 177:4281.
- Sabatos, C.A. et al. (2003) Nat. Immunol. 4:1102.
- Zhu, C. et al. (2005) Nat. Immunol. 6:1245.
- Koguchi, K. et al. (2006) J. Exp. Med. 203:1413.
- Frisancho-Kiss, S. et al. (2006) J. Immunol. 176:6411.
- Geng, H. et al. (2006) J. Immunol. 176:1411.
Product Datasheets
Citations for Human 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.
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Citations: Showing 1 - 10
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Phosphatidylserine binding directly regulates TIM-3 function
Authors: Courtney M. Smith, Alice Li, Nithya Krishnamurthy, Mark A. Lemmon
Biochemical Journal
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Integrated tumor stromal features of hepatocellular carcinoma reveals two distinct subtypes with prognostic/predictive significance
Authors: Wei Li, Jun Han, Kefei Yuan, Hong Wu
Aging (Albany NY)
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Immune microenvironment changes induced by neoadjuvant chemotherapy in triple-negative breast cancers: the MIMOSA-1 study
Authors: Sarradin V, Lusque A, Filleron T et al.
Breast cancer research : BCR
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Progressive immune dysfunction with advancing disease stage in renal cell carcinoma
Authors: David A. Braun, Kelly Street, Kelly P. Burke, David L. Cookmeyer, Thomas Denize, Christina B. Pedersen et al.
Cancer Cell
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BRAF Inhibition Is Associated with Enhanced Melanoma Antigen Expression and a More Favorable Tumor Microenvironment in Patients with Metastatic Melanoma
Authors: Dennie Tompers Frederick, Adriano Piris, Alexandria P. Cogdill, Zachary A. Cooper, Cecilia Lezcano, Cristina R. Ferrone et al.
Clinical Cancer Research
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Spatially resolved whole-transcriptomic and proteomic profiling of lung cancer and its immune-microenvironment according to PD-L1 expression
Authors: Jeon, YK;Koh, J;Lee, D;Kim, S;Song, SG;Han, B;Jeong, H;Kim, YA;Keam, B;Lee, SH;Kim, K;Chung, DH;
Cancer immunology research
Species: Human
Sample Types: Whole Tissue
Applications: Immunohistochemistry -
Circulating and intratumoral immune determinants of response to atezolizumab plus bevacizumab in patients with variant histology or sarcomatoid renal cell carcinoma
Authors: Saliby, RM;El Zarif, T;Bakouny, Z;Shah, V;Xie, W;Flippot, R;Denize, T;Kane, MH;Madsen, KN;Ficial, M;Hirsch, L;Wei, XX;Steinarter, JA;Harshman, LC;Vaishampayan, UN;Severgnini, M;McDermott, DF;Lee, GM;Xu, W;Van Allen, EM;McGregor, BA;Signoretti, S;Choueiri, TK;McKay, RR;Braun, DA;
Cancer immunology research
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Antiretroviral therapy duration and immunometabolic state determine efficacy of ex vivo dendritic cell-based treatment restoring functional HIV-specific CD8+ T cells in people living with HIV
Authors: M Calvet-Mir, I Sánchez-Ce, N Martín-Cóf, P Martínez-F, H de la Fuen, I Tsukalov, C Delgado-Ar, MJ Calzada, I de Los San, J Sanz, L García-Fra, F Sánchez-Ma, A Alfranca, MÁ Muñoz-Fern, MJ Buzón, E Martín-Gay
EBioMedicine, 2022-06-02;81(0):104090.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
High levels of Tim-3+Foxp3+Treg cells in the tumor microenvironment is a prognostic indicator of poor survival of diffuse large B cell lymphoma patients
Authors: W Zhong, X Liu, Z Zhu, Q Li, K Li
International immunopharmacology, 2021-04-14;96(0):107662.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Molecular Classification and Tumor Microenvironment Characterization of Gallbladder Cancer by Comprehensive Genomic and Transcriptomic Analysis
Authors: N Ebata, M Fujita, S Sasagawa, K Maejima, Y Okawa, Y Hatanaka, T Mitsuhashi, A Oosawa-Tat, H Tanaka, S Miyano, T Nakamura, S Hirano, H Nakagawa
Cancers, 2021-02-10;13(4):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Microtubule-Driven Stress Granule Dynamics Regulate Inhibitory Immune Checkpoint Expression in T Cells
Authors: DM Franchini, O Lanvin, M Tosolini, E Patras de, A Cammas, S Péricart, CM Scarlata, M Lebras, C Rossi, L Ligat, F Pont, PB Arimondo, C Laurent, M Ayyoub, F Despas, M Lapeyre-Me, S Millevoi, JJ Fournié
Cell Rep, 2019-01-02;26(1):94-107.e7.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Frontline Science: Tim-3-mediated dysfunctional engulfment of apoptotic cells in SLE
Authors: D Zhao, M Guo, B Liu, Q Lin, T Xie, Q Zhang, X Jia, Q Shu, X Liang, L Gao, C Ma
J. Leukoc. Biol., 2017-07-28;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Secretion of IFN-? Associated with Galectin-9 Production by Pleural Fluid Cells from a Patient with Extrapulmonary Tuberculosis
Authors: J Zhao, B Shiratori, H Chagan-Yas, M Matsumoto, T Niki, M Tanaka, Y Takahashi, O Usami, Y Ashino, T Hattori
Int J Mol Sci, 2017-06-28;18(7):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Tim-3 is a Marker of Plasmacytoid Dendritic Cell Dysfunction during HIV Infection and Is Associated with the Recruitment of IRF7 and p85 into Lysosomes and with the Submembrane Displacement of TLR9
Authors: JA Schwartz, KL Clayton, S Mujib, H Zhang, AK Rahman, J Liu, FY Yue, E Benko, C Kovacs, MA Ostrowski
J. Immunol, 2017-03-06;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: ICC -
Galectin-9 as a Predictive Marker for the Onset of Immune-Related Adverse Effects Associated with Anti-CCR4 MoAb Therapy in Patients with Adult T Cell Leukemia
Authors: TO Mohammed, H Chagan-Yas, Y Ashino, W Nakayama, Y Takahashi, T Shimomura, T Fujimoto, Y Watanabe, T Niki, H Suzushima, T Hattori
Tohoku J. Exp. Med, 2017-03-01;241(3):201-208.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
TIM-1 promotes hepatitis C virus cell attachment and infection
Authors: Guangxiang Luo
J. Virol., 2017-01-03;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Comprehensive analyses of tumor immunity: implications for cancer immunotherapy
Genome Biol, 2016-08-22;17(1):174.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Soluble T cell immunoglobulin mucin domain 3 is shed from CD8+ T cells by the sheddase ADAM10, is increased in plasma during untreated HIV infection, and correlates with HIV disease progression.
Authors: Clayton K, Douglas-Vail M, Nur-ur Rahman A, Medcalf K, Xie I, Chew G, Tandon R, Lanteri M, Norris P, Deeks S, Ndhlovu L, Ostrowski M
J Virol, 2015-01-21;89(7):3723-36.
Species: Human
Sample Types: Cell Culture Supernates
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Characterizing functional domains for TIM-mediated enveloped virus entry.
Authors: Moller-Tank S, Albritton L, Rennert P, Maury W
J Virol, 2014-04-02;88(12):6702-13.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
T cell Ig and mucin domain-containing protein 3 is recruited to the immune synapse, disrupts stable synapse formation, and associates with receptor phosphatases.
Authors: Clayton K, Haaland M, Douglas-Vail M, Mujib S, Chew G, Ndhlovu L, Ostrowski M
J Immunol, 2013-12-13;192(2):782-91.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: ICC, Immunoprecipitation, Western Blot -
Decreased galectin-9 and increased Tim-3 expression are related to poor prognosis in gastric cancer.
Authors: Jiang, Jing, Jin, Mei-Shan, Kong, Fei, Cao, Donghui, Ma, Hong-Xi, Jia, Zhifang, Wang, Yin-Ping, Suo, Jian, Cao, Xueyuan
PLoS ONE, 2013-12-10;8(12):e81799.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Tim-3 expression defines regulatory T cells in human tumors.
Authors: Yan J, Zhang Y, Zhang J, Liang J, Li L, Zheng L
PLoS ONE, 2013-03-05;8(3):e58006.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Tim-3 negatively regulates IL-12 expression by monocytes in HCV infection.
Authors: Zhang Y, Ma CJ, Wang JM, Ji XJ, Wu XY, Jia ZS, Moorman JP, Yao ZQ
PLoS ONE, 2011-05-26;6(5):e19664.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Dysregulation of TIM-3-Galectin-9 Pathway in the Cystic Fibrosis Airways.
Authors: Vega-Carrascal I, Reeves EP, Niki T, Arikawa T, McNally P, O'Neill SJ, Hirashima M, McElvaney NG
J. Immunol., 2011-01-24;0(0):.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: ICC, Immunoprecipitation, Western Blot -
Beneficial effect of galectin 9 on rheumatoid arthritis by induction of apoptosis of synovial fibroblasts.
Authors: Seki M, Sakata KM, Oomizu S, Arikawa T, Sakata A, Ueno M, Nobumoto A, Niki T, Saita N, Ito K, Dai SY, Katoh S, Nishi N, Tsukano M, Ishikawa K, Yamauchi A, Kuchroo V, Hirashima M
Arthritis Rheum., 2007-12-01;56(12):3968-76.
Species: Human
Sample Types: Whole Cells
Applications: ICC, Neutralization -
Addition of anti-TIM3 or anti-TIGIT Antibodies to anti-PD1 Blockade Augments Human T cell Adoptive Cell Transfer
Authors: Marina Martinez, Soyeon Kim, Naomi St. Jean, Shaun O’Brien, Lurong Lian, Jing Sun et al.
OncoImmunology
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Existence of intratumoral tertiary lymphoid structures is associated with immune cells infiltration and predicts better prognosis in early-stage hepatocellular carcinoma
Authors: Hui Li, Jinju Wang, Hailing Liu, Tian Lan, Lin Xu, Genshu Wang et al.
Aging (Albany NY)
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Integrative clinical and molecular characterization of translocation renal cell carcinoma
Authors: Ziad Bakouny, Ananthan Sadagopan, Praful Ravi, Nebiyou Y. Metaferia, Jiao Li, Shatha AbuHammad et al.
Cell Reports
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Transforming growth factor beta -induced epithelial-to-mesenchymal signature predicts metastasis-free survival in non-small cell lung cancer
Authors: Edna Gordian, Eric A. Welsh, Nicholas Gimbrone, Erin M. Siegel, David Shibata, Ben C. Creelan et al.
Oncotarget
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A distinct innate immune signature marks progression from mild to severe COVID-19
Authors: Stéphane Chevrier, Yves Zurbuchen, Carlo Cervia, Sarah Adamo, Miro E. Raeber, Natalie de Souza et al.
Cell Reports Medicine
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