63-3109-42

antibody from Invitrogen Antibodies

Targeting: HAVCR2 CD366, FLJ14428, Tim-3, TIM3, TIMD3

Provider product page for 63-3109-42

Flow cytometry

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Antibody data

Antibody Data
Antigen structure
References [3]
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Validations
Flow cytometry [1]
Other assay [4]

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Product number: 63-3109-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD366 (TIM3) Monoclonal Antibody (F38-2E2), Super Bright™ 600, eBioscience™
Antibody type: Monoclonal
Antigen: Other
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.
Antibody clone number: F38-2E2
Concentration: 5 µL/Test

HAVCR2 protein structure - 63-3109-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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.

Supportive validation

Submitted by: 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.

Supportive validation

Submitted by: 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

Supportive validation

Submitted by: 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