Our murine IFN-γ ELISpot kits (R&D Systems) were used according to the manufacturer’s suggested protocol. Freshly isolated splenocytes from naïve or tumor-bearing mice treated or untreated with anti-PD1 were resuspended in HL-1 serum-free medium (BioWhittaker) supplemented with 1% penicillin/streptomycin/l -glutamine, and were plated at 106 per well in triplicate. Splenocytes were restimulated with the following synthetic HY-specific peptides: (i) Uty: the immunodominant Db–restricted HY-derived peptide corresponding to amino acids 245 to 253 (WMHHNMDLI) of the Uty gene product; (ii) Dby: the immunodominant I-Ab–restricted (NAGFNSNRANSSRSS) HY-derived peptide; and as specific controls (iii) for Db binding: the E7 peptide (RAHYNIVTF) derived from HPV and (4 (link)) I-Ab peptide: the T. cruzi peptide (SHNFTLVASVIIEEA). After wash steps and incubation with a biotinylated IFN-γ detection antibody, alkaline-phosphatase conjugated to streptavidin was added to enumerate IFN-γ–positive spots.
Ifn γ elispot kits
Manufactured by R&D Systems
Sourced in United States
The IFN-γ ELISpot kits from R&D Systems are designed to detect and quantify interferon-gamma (IFN-γ) secreting cells. The kits utilize an enzyme-linked immunospot (ELISpot) assay principle to enumerate antigen-specific T cells or other IFN-γ producing cells.
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Lab products found in correlation
2 protocols using ifn γ elispot kits
1
Murine IFN-γ ELISpot Assay Protocol
2
ELISpot Analysis of Cytokine Secretion
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Cell suspensions from each group were prepared and stimulated as earlier described. The number of IL-4 and IFN-γ secreting cells were determined using mouse IL-4 and IFN-γ ELISpot kits (R&D; Minneapolis, USA), according to the manufacturer’s protocols. Spot forming units (SFU) were counted using the ELISpot ImmunoSpot S5 Analyzer (C.T.L., Germany) and analyzed using the C.T.L. ImmunoSpot image software version 5.1. The results were expressed as SFU for 1 × 106 cells.
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