Mouse ifn γ elisa kit

The secretion of interleukin‐2 (IL‐2), interleukin‐10 (IL‐10), tumor necrosis factor‐α (TNF‐α), and interferon‐γ (IFN‐γ) were assessed by ELISA kits. The human IL‐2 ELISA Kit (Abcam, ab174444), human IL‐10 ELISA Kit (Abcam, ab46034), human TNF‐α ELISA Kit (Abcam, ab181421), and human IFN‐γ ELISA Kit (Abcam, ab46025) were used to detect the concentrations in the cell supernatant. The mouse IL‐2 ELISA Kit (Abcam, ab100716), mouse TNF‐α ELISA Kit (Abcam, ab100747), and mouse IFN‐γ ELISA Kit (Abcam, ab100689) were used to determine the concentrations in the mouse serum samples. The absorbance (OD) was measured at a wavelength of 450 nm.

According to previous reports, RAW264.7 cells were seeded in 96-well plates at a density of 1 × 10⁴ cells/well and incubated in the above medium with 100 μM different DP of COS (chitobiose, chitotriose, chitotetraose, chitopentaose, chitohexaose, chitoheptaose, and chitooctaose) for 24 h and stimulated with LPS (200 ng/mL, 6 h). RAW264.7 cells were washed 3 times with cold PBS and centrifuged at 1,000 g for 10 min at 4°C. The cell pellet thus obtained was resuspended in 0.5 mL Tris buffer (20 mM, pH 7.5, 1 μg/mL chymostatin, 2 μg/mL of pepstatin A, 100 μM phenylmethylsulfonyl fluoride, and 5 μg/mL aprotinin). The cells were lysed by two freeze-thaw cycles [20 (link)]. The supernatants were collected via centrifugation at 15,000 g for 10 min at 4°C. The inflammatory production was measured by using the kits (mouse IL-1β ELISA Kit, ab100704; mouse IL-10 ELISA Kit, ab108870; mouse IL-17A ELISA Kit, ab100702; and mouse IFN-γ ELISA Kit, ab100689; Abcam, USA).

Splenocytes isolated from BALB/c mice were stimulated with anti-CD3 or anti-CD3 + IL-6 + TGF-β with or without 1.25–10 nM triptolide treatment for 4 days (primary culture). The supernatants were assayed for IL-17 using a mouse IL-17 ELISA kit (ab100702, Abcam).
Serum concentrations of IL-17 and interferon (IFN)-γ were detected using the mouse IL-17 ELISA kit (ab100702, Abcam) and a mouse IFN-γ ELISA kit (ab252363, Abcam), respectively.