IPEC-J2 monolayers were grown to confluence on 12-mm diameter, 0.4-μm pore size polycarbonate filters. A subset of monolayers was infected with C parvum, and the permeability of uninfected control and C parvum–infected monolayers was measured at 24 and 48 hours after infection by concurrent transepithelial passage of 3 permeability probes: creatinine, fluorescein isothiocyanate–4 kilodalton dextran, and rhodamine–70 kilodalton dextran (catalog C4255, 46944, and R9379; Millipore Sigma). Probes were prepared in cell culture media at a final concentration of 11 mg/mL. At the beginning of each flux period, 100 μL of the flux solution was added to the apical side of each insert to give a final volume and concentration of 500 μL and 2.2 mg/mL, respectively. After 2 hours of incubation, basolateral recovery of each probe was measured in a plate reader (Synergy HT; BioTek, Winooski, VT) using freshly prepared standards. Fluorescence of fluorescein and rhodamine B were measured at 490 and 555 nm using emission wavelengths of 520 and 585 nm, respectively. Creatinine was measured separately using a colorimetric assay (catalog MAK080; Millipore Sigma).
Mak080
Manufactured by Merck Group
Sourced in United States
MAK080 is a laboratory instrument designed for the measurement and analysis of various samples. It is capable of performing specific analytical functions, but a detailed description cannot be provided while maintaining an unbiased and factual approach without making interpretations or extrapolations.
Automatically generated - may contain errors
Lab products found in correlation
Mak055, by Merck Group (9 mentions)
Mak052, by Merck Group (6 mentions)
Mak006, by Merck Group (5 mentions)
Ag723 m, by Merck Group (3 mentions)
Blyscan sulfated glycosaminoglycan assay kit, by Biocolor (2 mentions)
Synergy ht, by Agilent Technologies (1 mentions)
R9379, by Merck Group (1 mentions)
Synergy mx, by Agilent Technologies (1 mentions)
Urea assay kit, by Merck Group (1 mentions)
Colorimetric detection kit, by Thermo Fisher Scientific (1 mentions)
210 infinite series, by Tecan (1 mentions)
Vivaspin, by Cytiva (1 mentions)
Creatinine assay kit, by Merck Group (1 mentions)
Vivacon 500, by Sartorius (1 mentions)
Mak066, by Merck Group (1 mentions)
A113 2, by Nanjing Jiancheng (1 mentions)
A111 2, by Nanjing Jiancheng (1 mentions)
Mak077, by Merck Group (1 mentions)
Mak116, by Merck Group (1 mentions)
19 protocols using mak080
1
Permeability Assay for C. parvum Infection
2
Creatinine-Normalized Molecule Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All molecule abundances were additionally normalized to creatinine concentrations of corresponding urine samples using a creatinine colorimetric assay kit (MilliporeSigma, catalog MAK080). Absorbance at 570 nm was measured using a Spectramax M2 microplate reader. Each sample was measured in duplicates, and all measured creatinine concentrations were within range of the standard curve.
3
Serum Creatinine and AST Analysis
4
Serum Creatinine and AST Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5
Creatinine Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Urine creatinine was measured using creatinine assay kit (MAK080; Sigma-Aldrich Corp., St Louis, MO) according to the manufacturer's instructions. Briefly, samples were passed through 10 KDa cut off filter to sieve out unwanted proteins. The urine creatinine concentration was measured by a colorimetric assay at 530 nm using a microplate reader (Synergy MX; Biotek Instruments Inc., VT). Serum creatinine concentration was measured with a serum-specific assay (700460; Cayman Chemical, Ann Arbor, MI), according to the manufacturer's protocol. This kit uses Jaffe's reaction, where picric acid in an alkaline condition reacts with creatinine to give a concentration dependent color.
6
Hematological and Biochemical Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Red blood cell (RBC) counts were established as specified by Brown (1976). Hemoglobin (Hb) concentration was quantified according to [17 ]. The serum concentrations of creatinine, urea alanine transaminase (ALT) and aspartate transaminase (AST) were calculated using standard kits (Sigma-Aldrich; MAK052, MAK055, MAK080 and MAK006, respectively). C-reactive protein (CRP) levels were evaluated according to the guidelines of a commercial kit (AG723-M; Sigma-Aldrich).
7
Serum Biomarker Quantification in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse blood samples were collected at the end of experiments. Serum urea, urea nitrogen (BUN), and creatinine were respectively assessed by Urea assay kit (MAK006, Sigma-Aldrich, St. Louis, MO, USA), Urea Nitrogen (BUN) colorimetric detection kit (EIABUN, Thermo Fisher Scientific, Waltham, MA), and Creatinine assay kit (MAK080, Sigma-Aldrich) according to the manufacturer’s instructions.
8
Urine Creatinine Normalization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A random subset of urine samples from the discover and validation cohort were selected and urine molecule abundance was compared prior to and after creatinine normalization. Urine creatinine concentration was measured using a creatinine colorimetric assay kit (Sigma Aldrich Catalog number MAK080). Absorbance was measured using a Spectramax M2 microplate reader at 570 nm (fluorometric (λex = 535/λem = 587 nm). If the creatinine concentration was out of the measurable range, the sample was excluded from the analysis. Creatinine measurement was done in triplicate for each urine sample. Statistical analysis was performed as described in statistical methods section. A random sample of 66 participants from the discovery (Haitian) cohort and 88 from the validation (Vietnam) cohort was analyzed before and after creatinine normalization. There was no difference in overall metabolite abundance or ROC characteristics when comparing samples that were normalized to osmolality alone vs. those that were normalized to both osmolality and creatinine. (Supplementary Table 1).
9
Glycosaminoglycan Quantification in Tissues and Urine
10
Urinary ß-CTX Measurement Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cross-linked C-terminal telopeptides of type I collagen (ß-CTX) were measured by an enzyme-linked immunosorbent assay (Catalog Number: EH3989, Fine-Biotech), as previously reported [46 (link)]. Assay values were corrected for urinary dilution by urinary creatinine concentration. The intra- and interassay coefficients of variation were <8% and <10%, respectively. All samples were analyzed in duplicates in the same assay to prevent interassay variation. Urinary creatinine was determined by a standard colorimetric method (Catalog Number: MAK080, Sigma-Aldrich, St. Louis, MO, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!