TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

Novus Biologicals | Catalog # NBP3-11989

Recombinant Monoclonal Antibody
Novus Biologicals
100% Guarantee

Key Product Details

Species Reactivity

Mouse

Applications

Immunohistochemistry, Block/Neutralize, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # 2C12

Format

Azide and BSA Free
View all TIM-3 Primary Antibodies »

Product Specifications

Immunogen

This antibody was raised by immunising rats with TIM-3:Ig fusion protein emulsified in complete Freund's adjuvant.

Specificity

This antibody is specific for murine TIM-3, a Th1-specific cell surface protein. TIM-3 is a type I transmembrane protein and contains an immunoglobulin and a mucin-like domain in its extracellular portion and a tyrosine phosphorylation motif in its cytoplasmic portion. This antibody binds to the BALB/c allele of TIM-3 while reactivity to the C57Bl/6 allele is significantly weaker.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Scientific Data Images for TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

Immunocytochemistry/ Immunofluorescence: TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free [NBP3-11989]

Immunocytochemistry/ Immunofluorescence: TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free [NBP3-11989]

Immunocytochemistry/Immunofluorescence: TIM-3 Antibody (2C12) - Chimeric [NBP3-11989] - Immunofluorescence analysis of paraformaldehyde fixed RAW264.7 cells on Shi-fix(TM) coverslips stained with the chimeric rabbit IgG version of 2C12 (NBP3-11989) at 10 ug/ml for 1h followed by Alexa Fluor(R) 488 secondary antibody (2 ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom NBP3-11989, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody followed by staining with Alexa Fluor(R) 488 secondary antibody.
TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

ELISA: TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free [NBP3-11989] -

ELISA: TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free [NBP3-11989] - Binding curves of NBP3-11989 (blue line) and isotype control (NBP2-52638; red line) to an ELISA plate coated with mouse TIM-3-Fc fusion protein at a concentration of 5 ug/ml. A 3-fold serial dilution from 3000 to 0.0169 ng/ml was performed using NBP3-11989. For signal detection, anti-rabbit IgG1 HRP antibody was used.
TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

Flow Cytometry: TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free [NBP3-11989] -

Flow Cytometry: TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free [NBP3-11989] - Paraformaldehyde fixed mouse splenocytes permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (isotype control, black line) or NBP3-11989 (blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Applications for TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

Application
Recommended Usage

Block/Neutralize

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This chimeric rabbit antibody was made using the variable domain sequences of the original Rat IgG1 format, for improved compatibility with existing reagents, assays and techniques.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.

Background: TIM-3

T cell immunoglobulin and mucin-domain containing 3 (TIM-3) is a type I transmembrane protein that functions in suppressing immune cell responses and is considered a checkpoint receptor (1,2). TIM-3 is an inhibitory receptor that was first identified as a cell marker for interferon-gamma (IFN-gamma)-producing CD4+ T helper (Th1) and CD8+ T cytotoxic (Tc1) cells (1,2). TIM-3 is also expressed on other immune cell types: regulatory T cells (Treg), natural killer (NK) cells, macrophages, and dendritic cells (DCs), as well as leukemia stem cells (LSCs) (1-3). Human TIM-3 protein is 301 amino acids (aa) in length with a theoretical molecular weight (MW) of 33.4 kDa and shares ~63% aa sequence identity with mouse TIM-3 (2,4). The TIM-3 protein has an extracellular IgV domain, a mucin and stalk domain with O- and N-glycosylation sites, a transmembrane domain, and an intracellular tail with conserved tyrosine residues (2,3). Ligands for TIM-3 include soluble galectin-9, high-mobility group protein B1 (HMGB1), phosphatidylserine (PtdSer), and carcinoembyronic antigen-related cell adhesion molecule-1 (CEACAM-1) (1-3,5). Each of these ligands interact with different regions of the TIM-3 IgV domain. In the absence of ligand binding, the unphosphorylated intracellular tyrosine residues of TIM-3 are associated with HLA-B associated transcript 3 (Bat3), which recruits Lck and this Bat3-Lck complex preserves T cell signaling (1-3,5). When TIM-3 is engaged, an intracellular signaling cascade is initiated to inhibit immune cell activation. (1-3,5). Specifically, the tyrosine residues of TIM-3 are phosphorylated, Bat3 is released, and this results in suppression of immune responses (1-3, 5). Persistent, sustained TIM-3 signaling eventually results in T cell exhaustion (1-3,5).

Dysregulation of TIM-3 expression is associated with autoimmune diseases as shown by studies where inhibition of TIM-3 using blocking antibodies worsened disease progression in experimental autoimmune encephalomyelitis (EAE) models of multiple sclerosis (MS) (2,3,5). Conversely, high levels of TIM-3 have been shown during viral infection as well as in many cancer types where its increased expression may be an indicator of poor prognosis (2,3,5). TIM-3 has emerged as a potential cancer immunotherapy target as preclinical studies blocking TIM-3 results in increased anti-tumor immunity and prevents tumor growth (3,5). Studies have suggested combination therapy of TIM-3 blockade with blockade of other checkpoint inhibitors such as programmed death 1 (PD-1) or lymphocyte activation gene 3 (LAG-3) is more effective than TIM-3 blockade alone (3,5).

References

1. Acharya N, Sabatos-Peyton C, Anderson AC. Tim-3 finds its place in the cancer immunotherapy landscape. J Immunother Cancer. 2020; 8(1):e000911. https://doi.org/10.1136/jitc-2020-000911

2. Das M, Zhu C, Kuchroo VK. Tim-3 and its role in regulating anti-tumor immunity. Immunol Rev. 2017; 276(1):97-111. https://doi.org/10.1111/imr.12520

3. Joller N, Kuchroo VK. Tim-3, Lag-3, and TIGIT. Curr Top Microbiol Immunol. 2017; 410:127-156. https://doi.org/10.1007/82_2017_62

4. Uniprot (Q8TDQ0)

5. Wolf Y, Anderson AC, Kuchroo VK. TIM3 comes of age as an inhibitory receptor. Nat Rev Immunol. 2020; 20(3):173-185. https://doi.org/10.1038/s41577-019-0224-6

Long Name

T Cell Immunoglobulin Mucin-3

Alternate Names

CD366, HAVcr-2, HAVCR2, KIM-3, SPTCL, TIM3, TIMD3

Gene Symbol

HAVCR2

Additional TIM-3 Products

Product Documents for TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for TIM-3 Antibody (2C12) - Chimeric - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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