IFN‐γ in supernatants was quantified with human IFN‐γ High sensitivity ELISA kit (Abcam) according to the manufacturer's instructions.
Human ifn γ high sensitivity elisa kit
Manufactured by Abcam
Sourced in United Kingdom, United States
The Human IFN‐γ High sensitivity ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of IFN‐γ levels in human cell culture supernatants, serum, and plasma samples.
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4 protocols using human ifn γ high sensitivity elisa kit
1
Quantification of IFN-γ by ELISA
2
Evaluating IL-12 and IL-15 Cytokine Effects
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The functionality of the IL-12 and IL-15 moieties of SON-1210 was evaluated by measuring IFNγ production from both human and monkey PBMCs in vitro after prior stimulation of the cells with PHA-L for 72 hours, then IL-2 for 24 h. Washed cell aliquots were incubated with the SON-1210, IL-12 (Peprotech, Cat# 200-12), or media alone for 48h, and the level of IFNγ was determined by ELISA (Human IFNγ High Sensitivity ELISA Kit [Abcam, Cat# ab46048]; Monkey IFNγ ELISA PRO Kit [Mabtech, Cat# 3421M-1HP-1]) (Supplementary Material Section 1.6
3
Serum Cytokine Profiling Protocol
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Serum cytokines were secondary outcome measures of this study. Interferon gamma (IFN-γ) was measured with an IFN-γ High-Sensitivity Human ELISA Kit (Abcam plc-Cambridge Science Park, Cambridge, UK) following to the manufacturer's instructions. Interleukin (IL)-12 concentrations were analyzed with a High-Sensitivity Human IL-12 (P70) ELISA kit (Genway Biotech, Inc., San Diego, CA, USA) using a Victor × 5 2,030 multi-label reader (PerkinElmer, Hopkinton, MA, USA) at 450 nm. IL-6, IL-1β, and tumor necrosis factor (TNF)-α concentrations in serum were measured using the Bio-Plex™ Reagent Kit (Bio-Rad Laboratories, Hercules, CA, USA).
4
Cytokine Panel Profiling via Multiplex Assays
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The serum high-sensitivity CRP (hs-CRP) level was analyzed with a CRP kit (Roche, Mannheim, Germany), and turbidity was measured by a Hitachi 7600 autoanalyzer (Hitachi, Tokyo, Japan). IL-1β, IL-6, and TNF-α levels were assessed via antibodies specific for each cytokine in a Bio-Plex reagent kit (Bio-Rad Laboratories, Hercules, CA, USA), and the resulting color reaction was analyzed with a Luminex 200 (Luminex Co., Austin, Texas, USA). IL-2, IL-12, and IFN-γ levels were analyzed using a human IL-2 ELISA kit (Cusabio Biotech, Houston, TX, USA), a high-sensitivity human IL-12 (p70) ELISA kit (Genway Biotech, Inc., San Diego, CA, USA), and an IFN-γ high-sensitivity human ELISA kit (Abcam plc, Cambridge, UK), respectively; the resulting color reactions were measured using a Victor × 5 2030 multilabel plate reader (PerkinElmer, Inc., Hopkinton, MA, USA) at an absorbance of 450 nm.
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