Bhb colorimetric assay kit

After the mice had been humanely killed at the completion of flexiVent analysis, approximately 300 μL of blood was collected via cardiac puncture from the right ventricle by using a 25-g needle attached to a 1-mL syringe. The blood was transferred into serum separator tubes (Becton Dickinson, Franklin Lanes, NJ) and centrifuged, and the serum was kept frozen at −80°C. The serum was used to determine the concentration of BHB (BHB colorimetric assay kit, Cayman Chemical, Ann Arbor, Mich) and immunoglobulins.

Anti-HMGCS2 (ab137043), anti-p-STAT6 (ab263947), anti-STAT6 (ab217998), and anti-p-JAK2 antibodies (ab92552) were obtained from Abcam. Anti-JAK2 antibody (3230T) was purchased from Cell Signaling Technology. Anti-ACAT1 (16215-1-AP) and anti-BDH1 (15417-1-AP) were purchased from Proteintech. Anti-ZO-1 (GB111981), anti-Occludin (GB111401), anti-F4/80 (GB113373), anti-Brdu (GB12051), and anti-PCNA (GB12010) were purchased from Servicebio.
Dextran sulfate sodium (DSS) was obtained from MP Biomedicals. Recombinant mouse GM-CSF (96-315-03-20) and recombinant mouse IL-4 (96-214-14-20) were obtained from Peprotech. BHB (S1031) and AS1517499 (S8685) were purchased from Selleck.cn. Cell Counting Kit-8 (CCK8) was obtained from Dojindo Laboratories, Japan. Clodronate liposomes (40337ES08) were obtained from Yeasen. Lipopolysaccharides (297-473-0), BHB sodium (13613-65-5), vancomycin (1404-93-9), neomycin-sulfate (1405-10-3), metronidazole (443-48-1), and ampicillin (69-52-3) were obtained from Sigma-Aldrich. BHB Colorimetric Assay Kit (700190) was obtained from Cayman Chemical. BCA Protein Assay Kit (P0012S) were obtained from Beyotime Biotechnology. Gentle Cell Disassociation Reagent (07174), DMEM/F12 with 15 mM HEPES (36254), and IntestiCult organoid growth medium (06005) were purchased from StemCell Technologies. Matrigel (356231) was purchased from Corning.

Commercial colorimetric assay kits were used to determine muscle/liver/brain total antioxidant capacity (Antioxidant assay kit #709001; Cayman Chemical, Ann Arbor, MI, USA) and serum BHB levels (BHB colorimetric assay kit #700190; Cayman Chemical), respectively, according to manufacturer’s instructions. For total antioxidant capacity determination, approximately 100 mg of frozen muscle/brain/liver was homogenized in kit assay buffer and centrifuged according to manufacturer’s instructions, and supernatants were assayed. Given that brain tissue in 8 month-fed rats was devoted to mitochondrial assays and Western blotting, brain total antioxidant capacity determination was not performed because of lack of available tissue. Following assay execution, all plates were read in a UV-Vis microplate reader (BioTek Synergy H1 Multi-Mode Reader; BioTek, Winooski, VT, USA) at absorbance readings according to manufacturer’s recommendations.