Creatinine colorimetric assay kit

BUN levels were measured using Quantichrom Urea assay kit (BioAssay Systems). Urinary albumin-to-creatinine ratio (ACR) was assessed by mouse albumin ELISA kit (Bethyl Laboratories) and creatinine colorimetric assay kit (Cayman Chemical). Assays were performed according to manufacturer’s protocols and read using EnSpire multimode plate reader (Perkin Elmer). Unpaired two-tailed t-tests were performed using Prism (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; whereas P > 0.05 was considered not significant, ns).

Estrogens and estrogen metabolites were extracted from urine samples of NSCLC patients and analyzed at Leidos Biomedical Research, Inc., Frederick (without hydrolysis). Urine samples from healthy Caucasian and Chinese American women were processed and analyzed at the University of Minnesota, as described previously [28 (link)]. Estrogens and estrogen metabolites (12 species, excluding 16-epiE₃, 17-epiE₃, 16-ketoE₂, and 3-OME₁) were measured using a Thermo Electron Quantum Discovery Max Triple Quadrupole LC-MS² instrument, with β-glucuronide and sulfate conjugates hydrolyzed [28 (link), 29 (link)]. Quantitative analysis was conducted using Thermo Electron Xcalibur proprietary software. Each sample was measured in duplicate and the coefficient of variation was less than 15%. Urinary creatinine concentrations were determined using the Creatinine Colorimetric Assay Kit (Cayman Chemical).

Plasma levels of BUN and creatinine were quantified using a Urea Nitrogen Colorimetric Detection Kit (Thermo Fisher Scientific, Waltham, MA; cat. #EIABUN) and Creatinine Colorimetric Assay Kit (Cayman Chemical, Ann Arbor, MI; cat. #700460), respectively, according to the manufacturers’ instructions.

The degree of kidney damage in Fas^lpr mice was evaluated by measuring the urinary ACR. A total of 50 μL of urine was collected from 6-week-old pre-onset, 19-week-old dPBS-, and CL-MSC-treated Fas^lpr mice (treatment on the 16th, 17th, and 18th week) by bladder massage at 3 weeks post treatment (PT). Urinary albumin concentrations were detected using a mouse albumin ELISA kit (Abcam, Cambridge, UK). Creatinine levels were detected by Jaffe’s reaction using a creatinine colorimetric assay kit (Cayman, Ann Arbor, MI, USA). Both were performed according to the manufacturer’s instructions. ACR is the ratio of albumin concentration in milligrams to creatinine concentration in grams.

Ammonia was measured in random spot urine and kidney homogenates (20 mg/0.5 mL deionized water) using Cell Biolabs, Inc. colorimetric Ammonia Assay (MET-5086, San Diego, CA) according to the manufacturer’s protocol. Urinary ammonia was normalized to creatinine utilizing the Creatinine Colorimetric Assay Kit (500701, Cayman Chemicals, Ann Arbor, Michigan).