Mouse il 10 elisa kit

RPMI 1640(SH30809.01), TRYPSIN 0.25% Solution (SH30042.01), and PBS buffer (SH30256.01B) were purchased from HyClone; Penicillin-streptomycin solution (100 × , C0222), PE anti-mouse CD40(124609), APC anti-mouse CD86 (105011), PerCP/Cy5.5 anti-mouse CD80 (104721), and FITC anti-mouse CD11c(117305) were purchased from Beyotime; Murine M-CSF(315-02-10 μg) and IL-4(214-14-20 μg) were purchased from PEPROTECH; FBS (P30-3302) was purchased from PAN; Goat Anti-Mouse IgA alpha chain (HRP) (ab97235), Goat Anti-Mouse IgG1 heavy chain (HRP) (ab97240), and Goat Anti-Mouse IgG2a heavy chain (HRP) (ab97245) were purchased from Abcam; Mouse TNF-α ELISA kit (1217202), Mouse IL-12p70 Elisa kit (1211202), Mouse IL-10 ELISA kit (1211002), and Mouse IL-6 ELISA kit (1210602) were purchased from Dakewe; Lowry protein concentration assay kit (PC0030), DAB color development kit (DA1010), and KDO (2-keto-3-deoxyoctane, K2755) was purchased from Solarbio; Quant-iT™ PicoGreen® dsDNA Reagent (P7581) was purchased from Thermo (Invitrogen).

SF was provided by Shanghai Biochempartner Co., Ltd. (Shanghai, China). Injectable soya lecithin was provided by Shanghai Taiwan Pharmaceutical Co., Ltd. (Shanghai, China). Coumarin-6 (C6) was bought from Aladdin Chemical Co., Ltd. (Shanghai, RPC). DSPE-rhodamine B was purchased from Ruixi Biological Technology Co., Ltd (Xi’an, China). Methylthiazol tetrazolium (MTT) were purchased from Sigma-Aldrich (US). APC anti-mouse CD3, FITC anti-mouse CD4, PE anti-mouse CD8a, PE anti-mouse CD25, and Alexa Fluor® 647 anti-mouse FOXP3 were purchased from eBioscience. Alexa Fluor ® 488 anti-mouse CD86, PerCP/Cy5.5 anti-mouse F4/80, and APC anti-mouse CD206 were bought from eBioscience. Mouse IL-12p70 Elisa kit, Mouse TGF-β1 Elisa kit, Mouse IL-10 Elisa kit, and Mouse TNF-α ELISA kit were purchased from DAKEWE. All other reagents were of analytical grade and obtained commercially.

The concentrations of pro-inflammatory (IFN-gamma and IL-6) and regulatory (IL-10 and TGF-β) cytokines in the culture supernatants of BMDMs incubated with rSj-Cys and other controls or in the sera collected from mice 12 h after macrophage transfer were determined using specific ELISA detection kits (Mouse IFN-gamma ELISA Kit, Mouse IL-6 ELISA Kit and Mouse IL-10 ELISA Kit from Dakewe Biotech, China and Mouse TGF-beta 1 ELISA Kit from ABclonal, USA) according to the manufacturer’s procedures. In order to remove the exogenously added IL-10 in the culture medium, the culture supernatant in the group with added IL-10 was removed after 24 h of culture, and cells were washed with PBS 3 times. The culture was continued with medium without IL-10 for another 24 h before harvested for IL-10 measurement.