The mAbs APC anti-mouse CD49b (clone DX5) and Annexin V/7-AAD kit were obtained from BD PharMingen (San Diego, CA, USA). FITC anti-mouse CD3ε (clone 145-2C11), PE/CY7 anti-mouse CD19 (clone 1D3), and PE anti-mouse NK1.1 (clone PK136) were obtained from Sungene Biotech (Tianjin, China). PE/Cy7 anti-mouse CD69 (clone H1.2F3), PE/Cy7 anti-mouse NKp46 (clone 29A1.4), and FITC anti-mouse NKG2A/C/E (clone 20d5) were from eBioscience (San Diego, CA, USA). FITC anti-mouse granzyme B (clone GB11), PE anti-mouse NKG2D (clone CX5), PE anti-mouse perforin (clone S16009A), PE anti-mouse IFN-γ (clone XMG1.2), PE anti-mouse FasL (clone MFL3), PerCP/Cy5.5 anti-mouse TRAIL (clone N2B2), FITC anti-mouse Ly49A (clone YE1/48.10.6), and buffers for staining, fixation, and permeabilization, as well as CFSE Cell Division Tracker Kit, were from BioLegend (San Diego, CA, USA). Mouse IFN-γ ELISA kit was from Dakewe Biotech Co., Ltd. (Shenzhen, China). Recombinant murine IL-2, IL-12, and IL-15 were from PeproTech Inc. (Rockhill, NJ, USA). Poly(I:C) was from InvivoGen (San Diego, CA, USA). EasySep™ mouse NK cell isolation kit was from STEMCELL Technologies, Inc. (Cambridge, UK).
Mouse ifn γ elisa kit
Manufactured by Dakewe
Sourced in China
The Mouse IFN-γ ELISA Kit is a quantitative in vitro assay designed for the measurement of mouse interferon-gamma (IFN-γ) in cell culture supernatants, serum, and plasma samples.
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Lab products found in correlation
Mouse tnf α elisa kit, by Dakewe (2 mentions)
Cfse cell division tracker kit, by BioLegend (1 mentions)
Poly 1 c, by InvivoGen (1 mentions)
Annexin v 7 aad kit, by BD (1 mentions)
Easysep mouse nk cell isolation kit, by STEMCELL (1 mentions)
Mouse granzyme b elisa kit, by Abcam (1 mentions)
96 well plate, by Corning (1 mentions)
Mouse lymphocyte separation medium, by Dakewe (1 mentions)
7 protocols using mouse ifn γ elisa kit
1
Murine NK Cell Activation Assay
2
Modulating T Cell Cytotoxicity Assay
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A total of 5.0 × 103 B16-F10 cells and 5.0 × 104 stimulated CD8+ T cells were seeded into 96-well plates (Corning) and incubated overnight. Cocultured cells were treated with IgG control (20 μg/mL), FreeαPD1 & αPDL1, NPαPD1 & NPαPDL1 or imNAαPD1 & αPDL1, the concentration of αPD1 and αPDL1 was 10 μg/mL. At 24 h post-treatment, the supernatant was collected, and the modulators of T cell-mediated cytotoxicity in the supernatant were quantified via Mouse IFN-γ ELISA Kit (Dakewe Biotech), Mouse Granzyme B ELISA Kit (Abcam), and Mouse Perforin 1 ELISA Kit (Abbexa) according to the manufacturer’s protocol.
3
Quantifying Plasma IFN-γ in 4T1 Tumor Mice
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Plasma was collected from 4T1 tumor-bearing mice at the indicated time point. The concentrations of plasma IFN-γ were determined using a Mouse IFN-γ ELISA Kit (Dakewe Biotech Co., Ltd., Shenzhen, China) according to the manufacturer's instructions.
4
Cytotoxic T-cell Assay on GL261 Glioma
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GL261 cells were seeded in 96-well plates at 4000 cells per well in 0.1 ml medium. The next day, murine splenic lymphocytes were isolated using the Mouse Lymphocyte Separation Medium (7211011, Dakewe) according to the manufacturer’s instructions. Then the appropriate number of splenic lymphocytes was added in each well on top of the adhered GL261 cells. The ratio of splenic lymphocytes (effector cells) to GL261 cells in co-cultures was 20:1. Forty-eight hours post co-cultures, the production and secretion of IFN-γ or TNF-α in cell supernatants was measured with a mouse IFN-γ ELISA kit (1210002, Dakewe) or a mouse TNF-α ELISA kit (1217202, Dakewe), and the viability of the adhered GL261 cells was detected by MTT.
5
Evaluating Tumor-Immune Interactions
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4T1 cells were treated with PBS, HFn, Dox (2 µm ) or Dox@HFn (10 µm ) for 24 h. Then, 4T1 cells were co‐cultured with BMDC for 24 h. Subsequently, T cells and BMDC were cocultured at a 10:1 ratio for an additional 24 h. CD69 expression was detected by flow cytometry (BD Biosciences, USA). The supernatants obtained from the co‐culturing of T cells and DC for 24 h were used for cytokine analysis. The levels of IFN‐γ were measured using the Dakewe biotech Mouse IFN‐γ ELISA kit, following the manufacturer's instructions. Similarly, the levels of TNF‐α were determined using the Dakewe biotech Mouse TNF‐α ELISA kit, following the manufacturer's guidelines.
6
Epididymal Fat IFNγ Secretion
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Nfatc3-/-and WT control mice, both on HFD, were killed and epididymal fat pad was isolated immediately and were cultured at 37°C in a humidified incubator (5% CO 2 ) overnight. The supernatant IFNγ levels were measured with a mouse IFNγ ELISA kit from Dakewe Biotech Co.
7
Evaluation of IFN-γ and TGF-β in Tumor-Bearing Mice
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As described above [19 (link)], splenic cells from differently treated and non-treated tumor-bearing mice were incubated with MMC inactivated CT26 (alive but not proliferative) at a R:S ratio of 10:1. The supernatant of the mixed lymphocyte and tumor cell culture (MLTC) was collected on day 3, and the concentrations of IFN-γ were determined using the mouse IFN-γ ELISA kit (Dakewe Biotech, Shenzhen, China). Serum from treated and control mice were collected and detected via the TGF-β ELISA kit (NeoBioscience Ltd., Beijing, China).
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