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Human TIM-3 Antibody

Catalog #: AF2365
29 Citations 1 Review Datasheet / COA / SDS
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AF2365
AF2365-SP
TIM-3 in Human Tonsil Using Dual RNAscope®ISH and IHC.
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Citations (29)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human TIM-3 Antibody Summary

Species Reactivity
Human
Specificity
Detects human TIM-3 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant mouse (rm) TIM‑3 is observed and less than 1% cross-reactivity with recombinant human TIM-1, rmTIM-1, and rmTIM-2 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human TIM‑3
Ser22-Arg200
Accession # Q8TDQ0
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
0.1 µg/mL
Recombinant Human TIM‑3 Fc Chimera (Catalog # 2365-TM)
Flow Cytometry
0.25 µg/106 cells
Human peripheral blood monocytes
Immunohistochemistry
3-15 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Dual RNAscope ISH-IHC
3-25 µg/mL
See below
Knockout Validated
TIM‑3 is specifically detected in HDLM‑2 human Hodgkin's lymphoma parental cell line but is not detectable in TIM‑3 knockout HDLM‑2 human Hodgkin's lymphoma cell line.
 

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

Dual RNAscope ISH-IHC Compatible TIM-3 antibody in Human Tonsil Using Dual RNAscope<sup>®</sup>ISH and IHC. View Larger

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).

Immunohistochemistry TIM-3 antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

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.

Knockout Validated View Larger

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.

Immunohistochemistry Detection of Human TIM-3 by Immunohistochemistry View Larger

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.

Simple Western Detection of Human TIM-3 by Simple Western View Larger

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

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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 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).

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. Sanchez-Fueyo, A. et al. (2003) Nat. Immunol. 4:1093.
  6. Khademi, M. et al. (2004) J. Immunol. 172:7169.
  7. Wiener, Z. et al. (2007) J. Invest. Dermatol. 127:906.
  8. van de Weyer, P.S. et al. (2006) Biochem. Biophys. Res. Commun. 351:571.
  9. Gielen, A.W. et al. (2005) J. Neuroimmunol. 164:93.
  10. Oikawa, T. et al. (2006) J. Immunol. 177:4281.
  11. Sabatos, C.A. et al. (2003) Nat. Immunol. 4:1102.
  12. Zhu, C. et al. (2005) Nat. Immunol. 6:1245.
  13. Koguchi, K. et al. (2006) J. Exp. Med. 203:1413.
  14. Frisancho-Kiss, S. et al. (2006) J. Immunol. 176:6411.
  15. Geng, H. et al. (2006) J. Immunol. 176:1411.
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 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.

29 Citations: Showing 1 - 10
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  1. Phosphatidylserine binding directly regulates TIM-3 function
    Authors: Courtney M. Smith, Alice Li, Nithya Krishnamurthy, Mark A. Lemmon
    Biochemical Journal
  2. 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)
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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
  22. 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
  23. 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
  24. 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
  25. 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
  26. 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)
  27. 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
  28. 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
  29. 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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Human TIM-3 Antibody
By Anonymous on 10/17/2018
Application: WB Sample Tested: Jurkat human acute T cell leukemia cell line,K562 human chronic myelogenous leukemia cell line,RPMI 8226 human multiple myeloma cell line Species: Human
Western Blot Human TIM-3 Antibody AF2365