Mouse mmp 12 kit price picokine

BALF was collected from male C57BL/6 mice (WT and AIM^−/−) by washing the lungs with 1.0 mL saline 15 times. Subsequently, the cells in the BALF were incubated in RPMI 1640 with 10% fetal bovine serum and 100 U/mL penicillin/streptomycin in a 24-well plate for 1 h. After washing twice with PBS, the adherent cells were used as AMs [31 (link)]. CSE was prepared as previously described [31 (link)]. Briefly, the mainstream smoke of two cigarettes (Marlboro) was bubbled through 15 mL of culture media and filtered through a 0.22-µm filter (defined as 100% CSE). The CSE was diluted to a concentration of 1% using culture medium. Recombinant murine AIM protein was produced in-house as previously described [7 (link)]. The AMs were incubated with 1% CSE, and 1 µg/mL rAIM was added to those derived from AIM^−/− mice every 6 h. After incubation, the cells were harvested for RNA isolation at 6 and 12 h. The culture media were also collected, and MMP-12 was assessed using ELISA (Mouse MMP-12 Kit Price PicoKine; BOSTER Biological Technology).

The mice were intubated and subjected to bronchoalveolar lavage (BAL) with 0.6 mL normal saline three times through a tracheal cannula. Total cell counts in the BALF were calculated using a hemocytometer. Red blood cells were removed using the Red Blood Cell Lysing Buffer Hybri-Max (Sigma-Aldrich). Cytospun BALF was stained with Diff-Quik Reagent (Sysmex, Kobe, Japan), and differential cell counts were calculated by counting > 400 cells per mouse using a microscope. After performing the BAL, the lungs were inflated with the Tissue-Tek O.C.T. Compound (Sakura Finetek Japan, Tokyo, Japan) (50% vol/vol) diluted with PBS containing 10% sucrose and stored at -80 °C. Proteins in the BALF were assessed using enzyme-linked immunosorbent assay (ELISA): interleukin (IL)-33 (Mouse IL-33 DuoSet ELISA; R&D Systems, Minneapolis, CA, USA) and MMP-12 (Mouse MMP-12 Kit Price PicoKine; BOSTER Biological Technology, Pleasanton, CA, USA).