Human ifn γ elisa kit

Cells were seed into six-well plate and incubated with Ecoli or Bt. The culture medium was collected after incubation for 48 hours. The concentration of GZMB and IFN-γ in the supernatant of CD8+ cells were measured using Human Granzyme B ELISA Kit and Human IFN-γ ELISA Kit (Abcam, Cambridgeshire, UK) according to the manufacturer’s protocols. The absorbance was measured at 450 nm in a microplate reader. All the experiments were performed in triplicate.

Th-related cytokines in peripheral blood and gingival crevicular fluid samples were detected. The gingival crevicular fluid was collected as follows. After the supragingival plaque and dental calculi were removed, aseptic cotton was used to keep the moisture, and the surface was wiped dry using sterile cotton balls. The dental face was blown dry using a pneumatic gun. Then #30 absorbent paper was gently inserted into the periodontal pocket on the side of cheek until there was slight resistance. After 30 s, the paper was taken out and placed into sterile tubes. The same operation was performed at an interval of 30 s. If there was visible blood or saliva contamination, the paper was discarded, and the sample was re-taken after 30 s. Gingival crevicular fluid samples were cryopreserved. Then the levels of IL-2, IL-4, IL-10, IL-17, tumor necrosis factor-β (TNF-β) and IFN-γ were determined using human IL-2 ELISA kit (ab174444), human IL-4 ELISA kit (ab215089), human IL-10 ELISA kit (ab46034), human IL-17 ELISA kit (ab119535), human TNF-β ELISA kit (ab229202) and human IFN-γ ELISA kit (ab46025) (Abcam, USA).

The level of CAR-T-secreted cytokines including human IFN-γ (human IFN-γ ELISA Kit; ab46025, Abcam, MA, USA), human TNF-α (human TNF-α ELISA Kit; ab46087; Abcam, MA, USA), and human IL-2 (human IL-2 ELISA Kit; ab100566; Abcam, MA, USA) was measured in the supernatant of the cells via enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions. Briefly, CAR-Ts were co-cultured with the target cells at an E:T ratio of 6:1, and the supernatant was collected 24 hours after incubation.

Concentrations of IFN-γ and TNF-α in 16HBE cell culture supernatant and rat serum were respectively determined using human IFN-γ ELISA kit (ab46025), human TNF-α ELISA kit (ab181421), rat IFN-γ ELISA kit (ab46107) and rat TNF-α ELISA kit (ab46070) (all from Abcam) following the manufacturer’s protocols.

Interferon (IFN)-γ and TNF-α levels in serum samples were quantitatively measured using the Human IFN-γ ELISA Kit (Abcam; Cambridge, UK) and Human TNF-α ELISA Kit (Abcam), following the manufacturer’s instructions.