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The EH2IL2 is a laboratory equipment product from Thermo Fisher Scientific. It is designed to perform a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach. More information may be available from the manufacturer.

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Lab products found in correlation

Opteia, by BD (1 mentions) Synergy h4 hybrid multi mode microplate reader, by Agilent Technologies (1 mentions) Corresponding kits, by Thermo Fisher Scientific (1 mentions) Bms228, by Thermo Fisher Scientific (1 mentions) Khc3011, by Thermo Fisher Scientific (1 mentions) Spectramax id3 multi mode microplate reader, by Molecular Devices (1 mentions) Synergy h4 hybrid multi mode microplate reader, by Bioteck (1 mentions)

6 protocols using eh2il2

1

Quantifying IL-2 Secretion in Transfected 293 Cells

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Flp-In T-REx 293 cells were plated at a density of 1.5 × 106 cells in one well of six-well plates. Twenty-four hours after transfection with WT IL-2 or IL-2 constructs containing inserted poly(A) tracks, medium was collected. Medium was centrifuged at 1,500 rpm in a centrifuge at 4°C for 10 min, and cell supernatant was collected. The supernatant was used immediately or stored at −80°C. A solid-phase sandwich ELISA (Thermo Fisher Scientific, EH2IL2) was used to measure IL-2 concentration from cell supernatant according to the manufacturer’s protocol (EH2IL2_ELISA_Kit_PI.pdf">https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0014639_EH2IL2_ELISA_Kit_PI.pdf). Cell supernatant was diluted with complete cell medium at a ratio of 1:10 to be within the concentration of a range of the standard curve. Diluted cell supernatant was further diluted to obtain 1:2, 1:4, 1:8, and 1:16 dilutions using complete medium. Absorbance measurements were taken using the Synergy H4 hybrid multi-mode microplate reader (BioTek Instruments) at 450–550 nm. Concentrations were interpolated from standard curves created with GraphPad Prism statistical software.
Powell G., Pavlovic Djuranovic S, & Djuranovic S. (2021). Gene dosage effects of poly(A) track-engineered hypomorphs. Molecular Therapy. Nucleic Acids, 26, 865-878.
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2

Quantifying Interleukin-2 Secretion

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Flp-In T-REx 293 cells were plated at a density of 1.5x10^6 cells in one well of the six-well plates. Twenty-four hours after transfection with WT IL2 or IL2 constructs containing inserted polyA tracks media was collected. Media from was centrifuged for 1,500 rpm centrifuge at 4°C for 10 mins, and cell supernatant was collected. The supernatant was used immediately or stored at -80°C. The solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) (Thermo Fisher, EH2IL2) was used to measure IL2 concentration from cell supernatant according to the manufacturer's protocol(https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0014639_EH2IL2_ELISA_Kit_PI.pdf). Cell supernatant was diluted with complete cell media at a ratio of 1:10 to be within the concentration of a range of the standard curve. Diluted cell supernatant was further diluted to obtain 1:2,1:4,1:8, and 1:16 dilutions using complete media. Absorbance measurements were taken using the Synergy H4
Hybrid Multi-Mode Microplate Reader (Bioteck) at 450 minus 550 nm. Concentrations were interpolated from standard curves created with GraphPad Prism statistical software.
Powell G., Pavlovic‐Djuranovic S., & Djuranović S. (2021). Gene dosage effects of polyA track engineered hypomorphs.
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3

Quantifying Human Cytokine Levels

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Medium was collected and centrifuged at 500 g for 10 min to remove debris. Human IFN-γ (555142; BD OptEIA) and human IL-2 (EH2IL2; Thermo Fisher Scientific) ELISA was performed according to the manufacturers’ protocol. Plates were analyzed on a Biotek Synergy2 microplate reader (Biotek) at wavelengths of 450 nm and a background of 550 nm.
Chheda Z.S., Kohanbash G., Okada K., Jahan N., Sidney J., Pecoraro M., Yang X., Carrera D.A., Downey K.M., Shrivastav S., Liu S., Lin Y., Lagisetti C., Chuntova P., Watchmaker P.B., Mueller S., Pollack I.F., Rajalingam R., Carcaboso A.M., Mann M., Sette A., Garcia K.C., Hou Y, & Okada H. (2018). Novel and shared neoantigen derived from histone 3 variant H3.3K27M mutation for glioma T cell therapy. The Journal of Experimental Medicine, 215(1), 141-157.
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4

Exosomal PD-L1 and T-cell Cytokines

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PD‐L1 concentration on exosomes was analyzed by serial dilution, and the concentration of PD‐L1 in exosomes derived from cells or plasma was calculated based on ELISA analysis. The CD8+ T‐cell supernatant was collected to detect the contents of IFN‐γ (#BMS228), TNF‐α (#KHC3011), and interleukin (IL)‐2 (#EH2IL2) following the protocols of corresponding kits (Thermo Fisher Scientific).
Yang J., Chen J., Liang H, & Yu Y. (2022). Nasopharyngeal cancer cell‐derived exosomal PD‐L1 inhibits CD8+ T‐cell activity and promotes immune escape. Cancer Science, 113(9), 3044-3054.
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5

Investigating Immune Modulation

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HCT116 cells and Jurkat T cells were co-cultured in the presence or absence of additives (10 μg/ml anti-PD-1 mAb (eBioscience #16–9989) or 250 μM peptide Ar5Y_4) in complete RPMI-1640 medium for 24 hours. Jurkat T cells alone were used as the reference. Supernatants were harvested and assessed for IL-2 by ELISA (Thermo scientific #EH2IL2).
Li Q., Quan L., Lyu J., He Z., Wang X., Meng J., Zhao Z., Zhu L., Liu X, & Li H. (2016). Discovery of peptide inhibitors targeting human programmed death 1 (PD-1) receptor. Oncotarget, 7(40), 64967-64976.
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6

Jurkat Cell Stimulation and CsA Treatment

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Jurkat cells, Clone E6–1 (TIB-152™, ATCC®, Manassas, VA) were cultured in medium composed of Advanced RPMI 1640 + 10% FBS + 10 mM HEPES + 100 units/mL penicillin + 100 μg/mL streptomycin to a density of 3×105/ml. Stimulation of Jurkat cells was with Phorbol 12-myristate 13-acetate (PMA) and ionomycin, each dissolved in DMSO and then added to culture medium to a final concentration of 20 ng/mL for PMA and 1 μg/mL for ionomycin. Immediately after the stimulation, cells were subject to treatment with CA192-CsA or free CsA dissolved in 2.5 % v/v DMSO at CsA concentrations from 10 pM to 100 nM for 6 hr at 37°C. IL-2 concentration in culture medium was assessed using an ELISA kit (EH2IL-2, Thermo Fisher Scientific, Waltham, MA) on a SpectraMax iD3 Multi-Mode Microplate Reader (Molecular Devices, San Jose, CA) at 450 nm.
Guo H., Lee C., Shah M., Janga S.R., Edman M.C., Klinngam W., Hamm-Alvarez S.F, & Mackay J.A. (2018). A novel elastin-like polypeptide drug carrier for cyclosporine A improves tear flow in a mouse model of Sjögren’s Syndrome. Journal of controlled release : official journal of the Controlled Release Society, 292, 183-195.
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