Alt gpt reagent

Liver TG, serum TG and serum cholesterol were measured with Triglyceride Colorimetric Assay Kit (Cayman Chemical #10010303) and Cholesterol Fluorometric Assay Kit (Cayman Chemical #10007640), respectively, according to manufacturer protocols. Serum insulin and glucagon concentrations were determined by Mouse Insulin ELISA KIT (Invitrogen, EMMINS) and Glucagon Quantikine ELISA Kit (R&D Systems, DGCG0), respectively, following manufacturer protocols. Liver NADPH and NADP were measured by NADP/NADPH-Glo^™ Assays (Promega #G9081) according to manufacturer’s protocol. ALT and AST assays were performed with ALT(GPT) Reagent (Thermo Scientific^™, TR71121) and AST/GOT Reagent (Thermo Scientific^™, TR70121), respectively, according to the manufacturer’s protocol. ATP, Acetyl-CoA and G6P concentrations were determined by ATP assay kit (Abcam, ab83355), Acetyl-Coenzyme A Assay Kit (Sigma, MAK039) and PicoProbe^™ Glucose-6-Phosphate Fluorometric Assay Kit (Biovision, K687), respectively according to the manufacturers protocols. Glucose and lactate were measured with Glucose colorimetric assay kit (Cayman Chemical #10009582) and Lactate Colorimetric/Fluorometric Assay Kit (Biovision, K607), respectively, according to the manufacturer’s protocols.

Mouse IgA and IgG ELISA kits from eBioscience were used according to the manufacturer’s protocols. Human serum IgA and IgG were measured in the first cohort from Newcastle, as described previously^{13 (link)}, or in cohort two at the UCSD Medical Center Laboratory. ELISA for the determination of OVA-specific IgA (anti-OVA-IgA) in the co-culture experiment was performed by adding culture supernatant to the ovalbumin-coated wells, incubating at 4 °C overnight, and washing with phosphate buffered saline with Tween 20. Rat anti-mouse IgA antibody (RMA-1, Biolegend) was added and incubated at room temperature for 1 h; thereafter, anti-rat HRP-conjugated antibody was added for a further 30 min. After washing with phosphate buffered saline with Tween 20, 1× TMB was added and the reaction was stopped with 2 N H₂SO₄, followed by detection in a microplate reader at 450 nm. Mouse serum ALT was measured using ALT (GPT) reagent (Thermo Fisher Scientific). Triglyceride and total cholesterol contents were measured using a Triglyceride Colorimetric Assay Kit (Cayman Chemical) and a Cholesterol Fluorometric Assay kit (Cayman Chemical) according to each manufacturer’s instructions.

Plasma levels of ALT (a marker of liver injury) were determined using the ALT/GPT Reagent (Thermo Fisher Scientific, Waltham, MA) according to the manufacturer’s instructions.

Plasma ALT levels, as a measurement of liver injury, was determined with ALT/GPT reagent from Thermo Fisher Scientific (Waltham, MA) according the manufacturer’s instructions.