Human TIM-3 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB23652-100
MAB23652-SP
Detection  of Human TIM‑3 by Western Blot.
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Human TIM-3 Antibody Summary

Species Reactivity
Human
Specificity
Detects human TIM-3 in direct ELISAs.
Source
Recombinant Monoclonal Rabbit IgG Clone # 2321C
Purification
Protein A or G purified from cell culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human TIM-3
Met1-Arg200
Accession # Q8TDQ0
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
Immunohistochemistry
3-25 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.
 

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.

Data Examples

Western Blot Detection  of Human TIM‑3 by Western Blot. View Larger

Detection of Human TIM‑3 by Western Blot. Western blot shows lysates of human spleen tissue, human tonsil tissue, and human thymus tissue. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human TIM‑3 Monoclonal Antibody (Catalog # MAB23652) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). 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.

Flow Cytometry Detection of Tim-3 in Human PBMC by Flow Cytometry. View Larger

Detection of Tim-3 in Human PBMC by Flow Cytometry. Human PBMC were stained with (A) Rabbit Anti-Human Tim-3 Monoclonal Antibody (Catalog # MAB23652) or (B) Normal Rabbit IgG Control Antibody (Catalog # MAB1050) followed by PE-conjugated Goat anti-Rabbit IgG Secondary Antibody (Catalog # F0110) and Mouse anti-Human CD14 APC-conjugated Monoclonal Antibody (Catalog # FAB3832A). View our protocol for Staining Membrane-associated Proteins.

Immunohistochemistry TIM‑3  in Human Tonsil. View Larger

TIM‑3 in Human Tonsil. TIM‑3 was detected in immersion fixed paraffin-embedded sections of human tonsil using Rabbit Anti-Human TIM‑3 Monoclonal Antibody (Catalog # MAB23652) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). 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 cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

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 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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 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). NK cells appear to transcribe the highest amounts of Tim-3 among lymphocytes, and when Tim-3 was cross-linked with antibodies it suppressed NK cell-mediated cytotoxicity (6). 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), and regulatory T cell development (5). TIM-3 inhibits the antitumor efficacy of DNA vaccines and chemotherapy by binding to the damage-associated molecular pattern molecule, HMGB1 (13).

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. Ndhlovu, L. et al. (2012) Blood 119:3734.
  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. Chiba, S. et al. (2012) Nat. Immunol. 13:832.
Long Name
T Cell Immunoglobulin Mucin-3
Entrez Gene IDs
84868 (Human); 171285 (Mouse); 102141722 (Cynomolgus Monkey)
Alternate Names
CD366; FLJ14428; HAVCR2; HAVcr-2; hepatitis A virus cellular receptor 2; kidney injury molecule-3; 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; TIMD-3; TIMD3KIM-3

Product Datasheets

FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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