Human TIM-3 Antibody

Name: Human TIM-3 Antibody
Brand: R&D Systems
SKU: AF2365
Rating: 5 (1 reviews)

Catalog # AF2365

Citations (17)

(1)

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Key Product Details

Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human TIM‑3 Ser22-Arg200 Accession # Q8TDQ0
Applications:	CyTOF-ready, Dual RNAscope ISH-IHC, Flow Cytometry, Immunohistochemistry, Knockout Validated, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
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.

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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

Item	Value
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.

Human TIM-3 Antibody Scientific Data Examples

	Dual RNAscope ISH-IHC Compatible 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 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 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.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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.
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.

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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).

Item	Value
References	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.
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, KIM-3, SPTCL, T cell immunoglobulin mucin 3, T cell immunoglobulin mucin-3, T-cell immunoglobulin and mucin domain-containing protein 3, TIM 3, TIM-3, TIM3, TIM3 T-cell membrane protein 3, TIMD-3, TIMD3, TIMD3KIM-3, hepatitis A virus cellular receptor 2, kidney injury molecule-3

Citations for Human TIM-3 Antibody (17)

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Citations are publications that use Bio-Techne products. Selected citations for Human TIM-3 Antibody include:

Species: Human
Sample Types: Whole Tissue
Applications: IHC

N Ebata et al. (2021), Molecular Classification and Tumor Microenvironment Characterization of Gallbladder Cancer by Comprehensive Genomic and Transcriptomic Analysis Cancers, 13(4).
PMID: 33578820
Species: Human
Sample Types: Whole Tissue
Applications: IHC

W Zhong et al. (2021), 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 International immunopharmacology, 96(0):107662.
PMID: 33864956
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

DM Franchini et al. (2019), Microtubule-Driven Stress Granule Dynamics Regulate Inhibitory Immune Checkpoint Expression in T Cells Cell Rep, 26(1):94-107.e7.
PMID: 30605689
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

D Zhao et al. (2017), Frontline Science: Tim-3-mediated dysfunctional engulfment of apoptotic cells in SLE J. Leukoc. Biol., 0(0).
PMID: 28754800
Species: Human
Sample Types: Whole Cells
Applications: ICC

JA Schwartz et al. (2017), 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 J. Immunol, 0(0).
PMID: 28264968
Species: Human
Sample Types: Whole Tissue
Applications: IHC

J Zhao et al. (2017), Secretion of IFN-? Associated with Galectin-9 Production by Pleural Fluid Cells from a Patient with Extrapulmonary Tuberculosis Int J Mol Sci, 18(7).
PMID: 28657598
Species: Human
Sample Types: Whole Tissue
Applications: IHC

TO Mohammed et al. (2017), 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 Tohoku J. Exp. Med, 241(3):201-208.
PMID: 28321034
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

(2016), Comprehensive analyses of tumor immunity: implications for cancer immunotherapy Genome Biol, 17(1):174.
PMID: 27549193
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Guangxiang Luo (2016), TIM-1 promotes hepatitis C virus cell attachment and infection J. Virol., 0(0).
PMID: 27807228
Species: Human
Sample Types: Cell Culture Supernates

Clayton K et al. (2015), 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. J Virol, 89(7):3723-36.
PMID: 25609823
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation

Clayton K et al. (2014), T cell Ig and mucin domain-containing protein 3 is recruited to the immune synapse, disrupts stable synapse formation, and associates with receptor phosphatases. J Immunol, 192(2):782-91.
PMID: 24337741
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

Moller-Tank S et al. (2014), Characterizing functional domains for TIM-mediated enveloped virus entry. J Virol, 88(12):6702-13.
PMID: 24696470
Species: Human
Sample Types: Whole Tissue
Applications: IHC

Yan J et al. (2013), Tim-3 expression defines regulatory T cells in human tumors. PLoS ONE, 8(3):e58006.
PMID: 23526963
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P

Jiang et al. (2013), Decreased galectin-9 and increased Tim-3 expression are related to poor prognosis in gastric cancer. PLoS ONE, 8(12):e81799.
PMID: 24339967
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation

Vega-Carrascal I et al. (2011), Dysregulation of TIM-3-Galectin-9 Pathway in the Cystic Fibrosis Airways. J. Immunol., 0(0).
PMID: 21263071
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Zhang Y et al. (2011), Tim-3 negatively regulates IL-12 expression by monocytes in HCV infection. PLoS ONE, 6(5):e19664.
PMID: 21637332
Species: Human
Sample Types: Whole Cells
Applications: ICC

Seki M et al. (2007), Beneficial effect of galectin 9 on rheumatoid arthritis by induction of apoptosis of synovial fibroblasts. Arthritis Rheum., 56(12):3968-76.
PMID: 18050192

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Human TIM-3 Antibody

Name: Anonymous

Application: Western Blot

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

Verified Customer | Posted 10/17/2018

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Human TIM-3 Antibody

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Key Product Details

Complete Your Research

Complete Your Experiment

Applications

Human TIM-3 Antibody Scientific Data Examples

Dual RNAscope ISH-IHC Compatible

Immunohistochemistry

Knockout Validated

Formulation, Preparation and Storage

Reconstitution Calculator

Background: TIM-3

Citations for Human TIM-3 Antibody (17)

Customer Reviews (1)

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Human TIM-3 Antibody

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