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Anti tim 3 bv421

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Anti-TIM-3-BV421 is a fluorochrome-conjugated antibody that binds to the T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) on the surface of cells. TIM-3 is an inhibitory immune checkpoint receptor that regulates T cell function. The BV421 fluorochrome allows for the detection and analysis of TIM-3-expressing cells using flow cytometry.

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Lab products found in correlation

Anti tnf α efluor 450, by Thermo Fisher Scientific (2 mentions) Anti ifn γ fitc, by BioLegend (2 mentions) Anti cd8 bv605, by BioLegend (2 mentions) Aurora flow cytometer, by Cytek (2 mentions) Foxp3 transcription factor kit, by Thermo Fisher Scientific (2 mentions) Anti cd3 buv395, by BD (2 mentions) Anti cd161 pe dazzle 594, by BioLegend (2 mentions) Anti granzyme b alexa fluor 700, by BioLegend (2 mentions) Zombie ultraviolet fixable viability dye, by BioLegend (2 mentions) Anti cd4 buv496, by BD (2 mentions) Anti cd69 buv563, by BD (2 mentions) Anti cd25 bv650, by BD (2 mentions) Anti vα7.2 pe cy7, by BioLegend (2 mentions) Anti lag 3 bv786, by BioLegend (2 mentions) Lsrii flow cytometer, by BD (1 mentions) Anti cd8 v500, by BD (1 mentions) Anti cd28 fitc, by BD (1 mentions) Anti cd3 alexa 780, by Thermo Fisher Scientific (1 mentions) Anti cd27 pe cy7, by Thermo Fisher Scientific (1 mentions) Anti cd4 qdot655, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

TIM-3 (58 citations) Anti-TIM-3 (20 citations) TIM-3-BV421 (9 citations)

4 protocols using anti tim 3 bv421

1

MAIT Cell Activation and Cytokine Production

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We cultured 2,000–5,000 MAIT cells from MAIT cells expanded in RPMI 1640 with 10% FBS or 2% Phx with 10,000–25,000 THP-1 cells to serve as APCs in RPMI with 10% FBS without cytokines overnight. We then stimulated the coculture of MAIT cells and THP-1 with 10 MOIs of strain 1100–2 fixed E. coli for 20 h and blocked extracellular transport with brefeldin A for a total of 4 h. For extracellular staining, we used Zombie ultraviolet fixable viability dye (BioLegend), anti-CD3-BUV395 (BD Biosciences), anti-CD8-BV605 (BioLegend), anti-CD4-BUV496 (BD Biosciences), anti-Vα7.2-PE-Cy7 (BioLegend), anti-LAG-3-BV786 (BioLegend), anti-CD69-BUV563 (BD Biosciences), anti-CD161-PE-Dazzle-594 (BioLegend), anti-TIM-3-BV421 (BioLegend), anti-CD25-BV650 (BD Biosciences), and anti-human PE-MR1–5-OP-RU tetramer (NIH Tetramer Core Facility). The cells were fixed and permeabilized using Foxp3/transcription factor kit (eBioscience) and stained intracellularly with anti-granzyme-B Alexa Fluor 700 (BioLegend), anti-TNF-α eFluor 450 (Invitrogen), and anti-IFN-γ-FITC (BioLegend). Sample data were acquired with a five-laser Cytek Aurora flow cytometer (Cytek) and analyzed using FlowJo software version 10 (BD Biosciences).
Bonnet G., Tyagi S., Libchaber A, & Kramer F.R. (1999). Thermodynamic basis of the enhanced specificity of structured DNA probes. Proceedings of the National Academy of Sciences of the United States of America, 96(11), 6171-6176.
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2

PBMC IFNγ ELISPOT Assay with Peptide Stimulation

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PBMCs responding to both NAE and AE in an IFNγ ELISPOT assay were pulsed with the peptides at 10μM in the presence of anti-CD28 and anti-CD49d. Monensin and brefeldin A were added 1 hour after peptide stimulation. The cells were incubated for an additional 11h. Following incubation, cells were surface stained for 30min at 4°C with dead cell dye (Invitrogen), anti-CD3-Alexa 780 (eBioscience), anti-CD4-Qdot655 (Invitrogen), and anti-CD8-V500 (BD Pharmingen) in the following panels: (1) anti-TIGIT-Percp/CY5.5 (Biolegend), anti-CD160-Alexa488 (eBioscience), anti-PD1-Alexa700 (Biolegend), anti-TIM3-BV421 (Biolegend) and anti-LAG3-PECy7 (Biolegend) and (2) anti-CD28-FITC (BD Pharmingen), anti-CD27-PECy7, anti-CD38-v450 (eBioscience), anti-CD57- Percp/CY5.5 (Biolegend), and anti-CD69-Alexa700 (Biolegend). The cells were then permeabilized and stained with anti-IFNγ-PE at 4°C for 30 min. At least 106 total events were acquired on an LSR II flow cytometer (BD Immunocytometry Systems), and analyzed using FlowJo (version 9.6.4; TreeStar Inc.). The criteria of positivity is the same as defined above for tetramer staining [73 (link)].
Qin K., Boppana S., Du V.Y., Carlson J.M., Yue L., Dilernia D.A., Hunter E., Mailliard R.B., Mallal S.A., Bansal A, & Goepfert P.A. (2019). CD8 T cells targeting adapted epitopes in chronic HIV infection promote dendritic cell maturation and CD4 T cell trans-infection. PLoS Pathogens, 15(8), e1007970.
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3

MAIT Cell Activation and Cytokine Production

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We cultured 2,000–5,000 MAIT cells from MAIT cells expanded in RPMI 1640 with 10% FBS or 2% Phx with 10,000–25,000 THP-1 cells to serve as APCs in RPMI with 10% FBS without cytokines overnight. We then stimulated the coculture of MAIT cells and THP-1 with 10 MOIs of strain 1100–2 fixed E. coli for 20 h and blocked extracellular transport with brefeldin A for a total of 4 h. For extracellular staining, we used Zombie ultraviolet fixable viability dye (BioLegend), anti-CD3-BUV395 (BD Biosciences), anti-CD8-BV605 (BioLegend), anti-CD4-BUV496 (BD Biosciences), anti-Vα7.2-PE-Cy7 (BioLegend), anti-LAG-3-BV786 (BioLegend), anti-CD69-BUV563 (BD Biosciences), anti-CD161-PE-Dazzle-594 (BioLegend), anti-TIM-3-BV421 (BioLegend), anti-CD25-BV650 (BD Biosciences), and anti-human PE-MR1–5-OP-RU tetramer (NIH Tetramer Core Facility). The cells were fixed and permeabilized using Foxp3/transcription factor kit (eBioscience) and stained intracellularly with anti-granzyme-B Alexa Fluor 700 (BioLegend), anti-TNF-α eFluor 450 (Invitrogen), and anti-IFN-γ-FITC (BioLegend). Sample data were acquired with a five-laser Cytek Aurora flow cytometer (Cytek) and analyzed using FlowJo software version 10 (BD Biosciences).
Labuz D., Cacioppo J., Li K., Aubé J, & Leung D.T. (2023). Enhancing Mucosal-Associated Invariant T Cell Function and Expansion with Human Selective Serum. ImmunoHorizons, 7(1), 116-124.
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4

Multiparameter Flow Cytometry of NK Cells

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Flow cytometry of NK cells was performed on a BD LSRII flow cytometer (BD Biosciences, San Jose, CA) and analyzed in FlowJo 9.9 (TreeStar Inc., Ashland, OR). Staining of NK cells was performed with four panels, and the antibodies used were: anti-CD56-BV605, anti-NKG2D-APC, anti-CD107a-PE-Cy7, anti-GranzymeB-FITC, anti-perforin-APC, anti-NKp46-PE-Cy7, anti-NKp30-APC, anti-NKp44-PE, anti-2B4-FITC, anti-4-1BB-PerCP-Cy5.5, anti-CD95-BV421, anti-Tim3-BV421, anti-TRAIL-PE, anti-CD122-PerCP-Cy5.5, anti-CD122-BV510, anti-CD95L-PE, anti-Ki67-BV421, anti-CD40L-BV510, anti-PD-1-PE-Cy7; all were obtained from BioLegend (San Diego, CA). Anti-CD16-APC-H7, PD-L1-PE-Cy7, and CD11a-FITC, were obtained from BD Biosciences (San Jose, CA). Anti-NKG2A-PE was from R&D Systems (Minneapolis, MN), and anti-CD158a-PerCP-Cy5.5 from eBioscience.
Jochems C., Tritsch S.R., Pellom S.T., Su Z., Soon-Shiong P., Wong H.C., Gulley J.L, & Schlom J. (2017). Analyses of functions of an anti-PD-L1/TGFβR2 bispecific fusion protein (M7824). Oncotarget, 8(43), 75217-75231.
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