62-3109-42
antibody from Invitrogen Antibodies
Targeting: HAVCR2
CD366, FLJ14428, Tim-3, TIM3, TIMD3
Antibody data
- Antibody Data
- Antigen structure
- References [3]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [4]
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- Product number
- 62-3109-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD366 (TIM3) Monoclonal Antibody (F38-2E2), Super Bright™ 436, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This F38-2E2 monoclonal antibody reacts with human CD366, also known as T cell immunoglobulin and mucin domain-containing protein 3 (TIM3) or HAVCR2. This cell surface receptor is expressed on activated CD4+ T cell subsets (e.g. Th1, Th17, and Treg), CD8+ T cells, monocytes, dendritic cells, and mast cells. Due to alternative splicing, CD366 exists as membrane-bound and soluble forms. Galectin-9 has been identified as the ligand for CD366. In humans, this receptor negatively regulates CD4+ T cells, influencing the secretion of some Th1- and Th17-related cytokines. CD366 has also been implicated in tolerance, autoimmune disease (e.g. multiple sclerosis), and HIV infection. Applications Reported: This F38-2E2 antibody has been reported for use in flow cytometric analysis. Applications Tested: This F38-2E2 antibody has been pre-titrated and tested by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Super Bright 436 can be excited with the violet laser line (405 nm) and emits at 436 nm. We recommend using a 450/50 bandpass filter, or equivalent. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Excitation: 405 nm; Emission: 436 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- F38-2E2
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references TIGIT(+) TIM-3(+) NK cells are correlated with NK cell exhaustion and disease progression in patients with hepatitis B virus‑related hepatocellular carcinoma.
SLC1A1 mediated glutamine addiction and contributed to natural killer T-cell lymphoma progression with immunotherapeutic potential.
TIM-3 is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines.
Yu L, Liu X, Wang X, Yan F, Wang P, Jiang Y, Du J, Yang Z
Oncoimmunology 2021;10(1):1942673
Oncoimmunology 2021;10(1):1942673
SLC1A1 mediated glutamine addiction and contributed to natural killer T-cell lymphoma progression with immunotherapeutic potential.
Xiong J, Wang N, Zhong HJ, Cui BW, Cheng S, Sun R, Chen JY, Xu PP, Cai G, Wang L, Sun XJ, Huang JY, Zhao WL
EBioMedicine 2021 Oct;72:103614
EBioMedicine 2021 Oct;72:103614
TIM-3 is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines.
Hastings WD, Anderson DE, Kassam N, Koguchi K, Greenfield EA, Kent SC, Zheng XX, Strom TB, Hafler DA, Kuchroo VK
European journal of immunology 2009 Sep;39(9):2492-501
European journal of immunology 2009 Sep;39(9):2492-501
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of unstimulated (left) or Anti-Human CD3 and CD28 Functional Grade Purified (Product # 16-0037-81 and Product # 16-0289-81)-stimulated (right) normal human peripheral blood cells with Anti-Human CD4 FITC (Product # 11-0049-42) and 0.25 µg of Anti-Human CD366 (TIM3) Super Bright 436. Total viable cells, as determined by 7-AAD Viability Staining Solution (Product # 00-6993-50), were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 1 Characterization of human anti-TIM-3 mAb. (A) Three purified anti-TIM-3 mAb (1G5, 2E2, and 4A4) were tested for binding to recombinant human TIM-3-Ig or human CTLA-4 Ig fusion proteins by ELISA. Comparative binding to TIM-3-Ig and CTLA-4-Ig is shown. Closed squares: hTIM-3; open squares: hCTLA4. (B) Anti-TIM-3 mAb were tested for their ability to stain CHO cells transfected with human TIM-3. Shading represents staining of isotype control. (C) Binding affinities of human TIM-3-specific mAb. Immobilized full-length human TIM-3 Fc fusion protein was used to measure the binding kinetics of the 1G5, 2E2, and 4A4 mAb. Binding of all three antibodies to immobilized human TIM-1 Fc fusion protein was not above background binding. (D) Anti-TIM-3 mAb were tested for their ability to bind different domains of recombinant TIM-3 expressed by 293 cells. 1G5 and 2E2 stained cells transfected with the IgV domain of human TIM-3, indicating specificity for this portion of the molecule. Shading represents staining of isotype control. Data are representative of three independent experiments.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 3 Expression of TIM-3 by short-term stimulated ex vivo CD4 + T cells do not correlate with cytokine production. (A) Total CD4 + T cells were isolated from PBMC of a normal healthy donor by negative selection, and stimulated with PMA plus ionomycin for 12 h. Cells were stained with 2E2-PE, followed by permeabilization and intracellular cytokine staining for IFN-gamma, IL-17, or IL-4. Data are representative of three independent experiments. (B) Sorting strategy to isolate TIM-3 + CD4 + T cells. Human spleen cell suspensions were stained with anti-CD4-APC and anti-TIM-3-PE, and sorted with a FACS Aria (Becton-Dickinson). Gates for TIM-3 + cells were set with isotype control PE (left panel). (C) FACS-sorted CD4 + TIM-3 + and CD4 + TIM-3 - cells were stimulated in vitro with plate-bound anti-CD3 and anti-CD28. Proliferation and cytokine production were assessed 48 h post-stimulation by uptake of tritiated thymidine and cytokine bead array, respectively. Results for two normal healthy donors are shown, and are representative of two independent experiments.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure2. The co-expression of TIGIT and TIM-3 is elevated on NK cells of progression patients with HBV-HCC. a-b Percentages of TIGIT + TIM-3 + NK cells on total NK cells and NK cell subsets(CD56 bright NK cells,CD56 dim NK cells, and CD56 - NK cells) from HBV-HCC, HDs, CHB and HBV-LC patients by flow cytometry analysis. c Correlation analysis of TIGIT and TIM-3 on NK cells from patients with HBV-HCC. d-e Flow-cytometry analyses (d) of TIGIT and TIM-3 were performed on PBMCs collected from HBV-HCC patients. Representative plots (e) display the expression of TIGIT + NK cells, TIM-3 + NK cells and total TIGIT + TIM-3 + NK cells from patients with progression (n = 61) and no progression (n = 72). P values were calculated by using the Kruskal-Wallis nonparametric H test (a-c). P values were obtained by the unpaired t test (d-e). * P < .05, ** P < .01, *** P < .001, **** P < .0001
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 5 Asparaginase treatment increased NKTCL cell sensitivity to anti-PD-1 antibody. (a) SLC1A1 expression on NK-92 cells transfected with SLC1A1 vector or control vector (upper panel) and SNK-6 cells transfected with SLC1A1 shRNA or scramble (lower panel). (b) Ki-67 and TIM-3 positivity of CD3+/CD8+ T cells in PBMC co-cultured with NK-92 cells (upper panel) or SNK-6 cells (lower panel) transfected with indicated vectors or shRNAs in medium with or without extra glutamine (2mM). (c) PD-L1 mRNA expression in NK-92 cells transfected with SLC1A1 vector or control vector (upper panel) and SNK-6 cells transfected with SLC1A1 shRNA or scramble (lower panel) upon asparaginase (10 IU/mL) treatment. The control vector or scramble values were normalized to 1, respectively. (d and e) Median fluorescence intensity of PD-L1 (d) on NK-92 cells (upper panel) or SNK-6 cells (lower panel), as well as Ki-67 and TIM-3 positivity of CD3+/CD8+ T cells in PBMC co-cultured with NK-92 cells (upper panel) or SNK-6 cells (lower panel) upon indicated treatment. (f) Gene expression correlation of tumor SLC1A1 with PD-L1 in NKTCL patients (n=128). (g) Tumor EAAT3 expression according to the TSIM, HEA, and MB subtypes in NKTCL patients (n=100). (h) PD-L1 mRNA expression of NK-92 cells transfected with TP53 R248Q, TP53 R273H, EP300 vector, or MGA shRNA upon indicated treatment. (i) Ki-67 positivity of CD3+/CD8+ T cells in PBMC co-cultured with NK-92 cells transfected with TP53 R248Q, TP53 R273H, EP300 vec