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Anti lag3

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Anti-LAG3 is a laboratory reagent that can be used to detect the expression of the Lymphocyte-Activation Gene 3 (LAG3) protein. LAG3 is a surface receptor expressed on activated T cells and regulatory T cells, and it plays a role in the regulation of immune responses. The Anti-LAG3 reagent can be used in various immunological and cell-based assays to study the expression and function of LAG3.

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Lab products found in correlation

Anti pd 1, by BD (5 mentions) Anti tim3, by BD (4 mentions) Anti ccr7, by BD (3 mentions) Anti cd8, by BD (2 mentions) Lsrii flow cytometer, by BD (2 mentions) Anti cd3, by BD (2 mentions) Anti cd4, by BD (2 mentions) Anti ve cadherin, by Abcam (2 mentions) Anti vinculin, by Santa Cruz Biotechnology (2 mentions) Anti cd45ro, by BD (2 mentions) Anti cd45 apc, by BD (2 mentions) Anti fak, by Abcam (2 mentions) Texas red phalloidin, by Thermo Fisher Scientific (2 mentions) Efluor 670, by BD (2 mentions) Anti alcam pe, by Abcam (2 mentions) Anti icam1 pe, by Abcam (2 mentions) Anti vcam1 pe, by Abcam (2 mentions) Anti chs1, by Abcam (2 mentions) Anti hif 1, by BD (2 mentions) Anti pzap 70 fitc, by BD (2 mentions)

7 protocols using anti lag3

1

Multiparameter Flow Cytometry Analysis

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Anti-CD3, anti-CD4, anti-CD8, anti-CD183, anti-CD196, anti-CD25, anti-CD127, anti-PD1, and anti-LAG3 were from BD Biosciences (California, USA). Peripheral blood samples were surface-labeled with antibodies for 10 min at room temperature. Then, red blood cells in the blood were lysed with red blood cell lysate (BD Biosciences, California, USA). The cells were washed with PBS twice. The CBA (Cytometric Bead Array) test kit was purchased from BD Biosciences. The LSR-II Flow Cytometer (BD Biosciences, California, USA) was used for the flow cytometry experiment. The Kaluza software (Beckman) was used for data analysis.
Liu Y., Guo X., Zhan L., Wang L., Wang X, & Jiang M. (2021). LAG3 and PD1 Regulate CD8+ T Cell in Diffuse Large B-cell Lymphoma Patients. Computational and Mathematical Methods in Medicine, 2021, 4468140.
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2

Multiparametric Immune Profiling of T-cell Subsets

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MyLa or HuT78 cell pellets were resuspended in FACS buffer (PBS, 5% FBS) and stained with predetermined antibody volumes: anti-CD107a (Cat# 555800, BD Bioscience), anti-IFNγ (Cat# 506510, Biolegend), anti-PD1 (Cat# 560908, BD Biosciences), anti-PD-L1 (Cat# 563741, Biolegend), anti-ICOS (Cat# 564778, Biolegend), anti-CTLA4 (Cat# 561717, BD Biosciences), anti-TIM3 (Cat# 565566, BD Biosciences), anti-LAG3 (Cat# 369318, BD Biosciences) and the fixation/permeabilization solution kit (Cat# 554714, BD Bioscience). Cells were analyzed using a Fortessa flow cytometer (BD Bioscience, San Jose, CA). The FlowJo 10.6.2 software program (Tree Star, Inc.) was used for data analysis.
Han Z., Estephan R.J., Wu X., Su C., Yuan Y.C., Qin H., Kil S.H., Morales C., Schmolze D., Sanchez J.F., Tian L., Yu J., Kortylewski M., Rosen S.T, & Querfeld C. (2021). MiRNA regulation of T cell exhaustion in cutaneous T cell lymphoma. The Journal of investigative dermatology, 142(3 Pt A), 603-612.e7.
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3

Multiparameter Flow Cytometry Analysis

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Fluorochrome-conjugated isotype controls, anti-CD123, anti-CD3, anti-CD4, anti-CD8, anti-CD33, anti-CD45, anti-CD45RA, anti-CD62L, anti-CCR7, anti-PD-1, anti-TIM3, anti-LAG3, anti-Annexin-V, 7-AAD, DAPI, and anti-mouse CD45 were purchased from BD Biosciences or BioLegend (San Diego, CA). Analysis was performed on at least 20,000 cells per sample using a LSR II flow cytometer (BD Biosciences) and a Gallios Flow Cytometer, and analyzed using Kaluza Analysis Software (Beckman Coulter, Indianapolis, IN).
Mu-Mosley H., Ostermann L., Muftuoglu M., Vaidya A., Bonifant C.L., Velasquez M.P., Gottschalk S, & Andreeff M. (2022). Transgenic Expression of IL15 Retains CD123-Redirected T Cells in a Less Differentiated State Resulting in Improved Anti-AML Activity in Autologous AML PDX Models. Frontiers in Immunology, 13, 880108.
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4

Multimodal Analysis of Cellular Adhesion Proteins

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Anti-FAK, anti-ALCAM-PE, anti-ICAM1-PE, anti-VCAM1-PE, anti-VE cadherin, anti-von Willebrand (vWF) factor, anti-CHS1 were purchased from Abcam (Cambridge, MA, USA). OX124, mouse anti-human monoclonal antibody against HS, was a kind gift from Dr. Marion Brown, Oxford, UK. Biotin-labeled Anti-CD18 KIM127 was purchased from Exploratory Research Cell Tech Therapeutics Ltd. (Slough, UK). Anti-phospho-SLP-76; pTyr128, anti-VE-cadherin were purchased from Cell signaling Technologies (Danvers, MA, USA). Anti-HIF-1, anti-pZAP-70-FITC, anti-pZAP-70-PE, anti- LAG3, anti-TIM3, anti-PD-1, eFluor 670, anti-CD45-APC, anti-CHS-PerCP, anti-CCR7, anti-CD45RO were purchased from BD biosciences (Franklin Lakes, NJ, USA). Anti-Talin-1 and anti-Vinculin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat anti-human Fc-PE was purchased from Millipore (Billerica, MA, USA). Alexa Fluor (AF) labelled secondary anti-mouse, anti-goat anti-rabbit antibodies, streptavidin AF 488 antibody; Texas Red Phalloidin and AF488 Phalloidin were purchased from Life Technologies (Carlsbad, CA, USA). Anti-Fab DyLight 488 was purchased from Jackson ImmunoResearch (Suffolk, UK)
Samaha H., Pignata A., Fousek K., Ren J., Lam F., Stossi F., Dubrulle J., Salsman V.S., Krishnan S., Hong S.H., Baker M.L., Shree A., Gad A.Z., Shum T., Fukumura D., Byrd T.T., Mukherjee M., Marrelli S.P., Orange J.S., Joseph S.K., Sorensen P.H., Taylor M.D., Hegde M., Mamonkin M., Jain R.K., El-Naggar S, & Ahmed N. (2018). A Homing System Targets Therapeutic T-cells to Brain Cancer. Nature, 561(7723), 331-337.
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5

Cytotoxicity Assay for Allogeneic T-cell Responses

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To measure cytotoxicity in the allogeneic assays, purified activated CD4+ or CD8+ T-cells from HLA-A*02:01-negative donors were incubated with HLA-A*02:01-positive PDAC cell lines at a ratio of 2:1. Where indicated, soluble LAG-3 (#2319-L3-050; R&D Systems), anti-LAG-3 (clone 17B4; Novus Biologicals; #NBP1-97657), anti-PD-L1 (clone 29E.2A3; #329716; BioLegend) or a combination of anti-LAG-3 and anti-PD-L1 antibodies or soluble LAG-3 and anti-PD-L1 were added at 10 μg/mL. After overnight incubation, the cells were stained with BV421-conjugated anti-HLA-A2 (#740082; BD), Calcein-AM and Ethidium Homodimer 1 (#L3224; Invitrogen) to determine cell death. Target PDAC cells were gated on HLA-A2 positive cells. The cytotoxicity of peptide-stimulated CD4+ T-cells on Capan-1 cells was determined through measurement of lactate dehydrogenase (LDH) released into the culture medium with the CytoTox 96® Non-Radioactive Cytotoxicity Assay Kit (#G1780; Promega). The assay was carried out according to the manufacturer’s instructions. Results are expressed as relative cytotoxicity, that is, cell death of Capan-1 cells by peptide-stimulated CD4+ T-cells in relation to unstimulated CD4+ T-cells.
Baleeiro R.B., Bouwens C.J., Liu P., Di Gioia C., Dunmall L.S., Nagano A., Gangeswaran R., Chelala C., Kocher H.M., Lemoine N.R, & Wang Y. (2022). MHC class II molecules on pancreatic cancer cells indicate a potential for neo-antigen-based immunotherapy. Oncoimmunology, 11(1), 2080329.
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6

Antigen-Specific Suppressive Assay Protocol

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For the antigen-specific suppressive assay, 1 × 105 DO11.10 splenic CD4+CD25− T cells as responder cells were stimulated with γ-irradiated (3000 rad) and OVA323–339 peptide-pulsed cells. The suppressor cells were seeded with responder cells at a ratio of 1:1. [3H]-thymidine (1 μCi in each well, PerkinElmer, Boston, MA) was added after 3 days, and the cells were harvested after 16–18 hours. During the neutralization or blockade assay, cells were treated with the anti-CD16/32 antibody (101310, BioLegend, San Diego, CA) prior neutralizing antibody, including anti-IL-10 (554463, BD), anti-IL-10 receptor (55012, BD), anti-CTLA4 (106204, BioLegend), anti-LAG3 (552379, BD), and the relative isotype control antibodies. The results were presented in counts per minute (c.p.m.).
Chien C.H., Yu H.C., Chen S.Y, & Chiang B.L. (2017). Characterization of c-Maf+Foxp3− Regulatory T Cells Induced by Repeated Stimulation of Antigen-Presenting B Cells. Scientific Reports, 7, 46348.
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7

Multimodal Analysis of Cellular Adhesion Proteins

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Anti-FAK, anti-ALCAM-PE, anti-ICAM1-PE, anti-VCAM1-PE, anti-VE cadherin, anti-von Willebrand (vWF) factor, anti-CHS1 were purchased from Abcam (Cambridge, MA, USA). OX124, mouse anti-human monoclonal antibody against HS, was a kind gift from Dr. Marion Brown, Oxford, UK. Biotin-labeled Anti-CD18 KIM127 was purchased from Exploratory Research Cell Tech Therapeutics Ltd. (Slough, UK). Anti-phospho-SLP-76; pTyr128, anti-VE-cadherin were purchased from Cell signaling Technologies (Danvers, MA, USA). Anti-HIF-1, anti-pZAP-70-FITC, anti-pZAP-70-PE, anti- LAG3, anti-TIM3, anti-PD-1, eFluor 670, anti-CD45-APC, anti-CHS-PerCP, anti-CCR7, anti-CD45RO were purchased from BD biosciences (Franklin Lakes, NJ, USA). Anti-Talin-1 and anti-Vinculin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat anti-human Fc-PE was purchased from Millipore (Billerica, MA, USA). Alexa Fluor (AF) labelled secondary anti-mouse, anti-goat anti-rabbit antibodies, streptavidin AF 488 antibody; Texas Red Phalloidin and AF488 Phalloidin were purchased from Life Technologies (Carlsbad, CA, USA). Anti-Fab DyLight 488 was purchased from Jackson ImmunoResearch (Suffolk, UK)
Samaha H., Pignata A., Fousek K., Ren J., Lam F., Stossi F., Dubrulle J., Salsman V.S., Krishnan S., Hong S.H., Baker M.L., Shree A., Gad A.Z., Shum T., Fukumura D., Byrd T.T., Mukherjee M., Marrelli S.P., Orange J.S., Joseph S.K., Sorensen P.H., Taylor M.D., Hegde M., Mamonkin M., Jain R.K., El-Naggar S, & Ahmed N. (2018). A Homing System Targets Therapeutic T-cells to Brain Cancer. Nature, 561(7723), 331-337.
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