C57Bl/6 female mice were infected intraperitoneally with 103 freshly purified RH-GFP parasites in a volume of 200 µl phenol-red-free DMEM. Beginning 1 h post infection, mice received 10 mg/kg SB505124 or DMSO alone intraperitoneally once daily. The drug had no apparent effect on the health of the animals during this time course. On day 5 post-infection, mice were sacrificed by CO2 asphyxiation and PECs collected in a 5 ml PBS-lavage. Cells were washed, stained with anti-mouse APC-conjugated CD45, and analyzed by flow cytometry using a FACSCalibur cytometer (BD Biosciences, San Jose, CA). 105 events were counted and subsequent analysis was performed using Summit (Dako, Carpinteria, CA). IFNγ was measured using Mouse IFNγ ‘Femto-HS’ High Sensitivity ELISA (eBioscience; San Diego, CA).
Mouse ifnγ femto hs high sensitivity elisa
Manufactured by Thermo Fisher Scientific
The Mouse IFNγ 'Femto-HS' High Sensitivity ELISA is a laboratory equipment product designed to detect and measure the concentration of interferon-gamma (IFNγ) in mouse samples. It employs a highly sensitive enzyme-linked immunosorbent assay (ELISA) technique to quantify IFNγ levels with high accuracy.
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2 protocols using mouse ifnγ femto hs high sensitivity elisa
1
Murine Toxoplasma Infection and IFNγ Assay
2
Measuring Cytokine Secretion and sRAGE in Bone Marrow Cell Cultures
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We plated TIRAP- or vector-transduced bone marrow cells at a concentration of 2 × 106 cells/ml in serum-free DMEM supplemented with 1% BSA and cultured them overnight. We recovered the supernatants, and centrifuged (3,500 rpm, 5 min) and assayed them for Ifnγ, Il10, and Hmgb1. Serum from peripheral blood collected at end point from mice transplanted with TIRAP- or vector-transduced HSPCs was used to measure Ifnγ. Mouse IL-10 ELISA Ready-Set-Go!, Mouse IFNγ "Femto-HS" High Sensitivity ELISA, and HMGB1 ELISA kits from eBioscience were used according to manufacturer’s instructions. Similarly, overnight-conditioned medium was collected from 106 bone marrow cells transduced with TIRAP, TIRAP-2A-sRAGE, or vector control to perform ELISA to detect soluble RAGE (sRAGE) using the Human RAGE Quantikine ELISA kit as per the manufacturer’s instructions.
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