Tgf β elisa kits

TLR2 agonist Pam3CSK4 was synthesized from InvivoGen (San Diego, USA). PE/CY5.5-anti- Gr-1, PE-anti-CD11b antibodies, TNF-α, IL-6, IL-10, IL-1β and TGF-β ELISA kits were purchased from BioLegend Inc. (San Diego, USA); PE/CY5.5 Rat IgG2b (κ) and PE Rat IgG2b (κ) were used as isotype controls (BioLegend Inc, San Diego, USA). methicillin-resistant S. aureus (ATCC43300) was purchased from Wenzhou Kont Biology & Technology Co. LTD (Jiangsu, China); Fluorescein isothiocyanate (FITC) was purchased from Sigma (Saint Louis, USA); Neutrophil Isolation Kit was purchased from Miltenyi Biotec (Cologne, Germany); TRIzol solution, a reverse transcription-PCR (RT-PCR) Kit and fluorescence quantitative PCR Kit were purchased from Takara Bio Inc. (Tokyo, Japan). Primers were synthesized by Invitrogen Inc. (Shanghai, China). Neutrophils Oxidative Burst Quantitative Assay Kit and ELISA Kit for Lactoferrin were purchased from Absin Inc. (Shanghai, China) and Cloud-Clone Corp. (Houston, USA), respectively.

The effect of ginsenoside Rg3 on the expression levels of pro-inflammatory cytokines and anti-inflammatory cytokines induced by LPS were measured using mouse TNF-α, IL-1β and IL-6, interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) ELISA kits (BioLegend, United States) according to the manufacturer’s protocols. Fifty microliters of a serial dilution gradient standard or sample was added to the appropriate wells and incubated at room temperature for 2 h. Then, the plate was washed 4 times with 1 × wash buffer. Subsequently, 100 μL of detection antibody solution was added to each well and incubated at room temperature for 1 h. After washing the plate 4 times with 1 × wash buffer, 100 μL of avidin-HRP solution was added to each well and incubated at room temperature for 30 min. One hundred microliters of substrate solution and 100 μL of stop solution were added to each well in turn; finally, the OD value was determined at 450 nm using a microplate reader.

Mouse tumor tissues were homogenized with extraction buffer. The supernatant was analyzed using mouse IL-10 (Biolegend, 431417) or TGF-β ELISA kits (Biolegend, 433007) according to the manufacturer’s instructions.