Plasma Biomarker Measurement in Mice

Heparin plasma was prepared from venous blood collected from the right ventricle of anesthetized mice. Mice were fasted at least 5 hours before blood collection. Alkaline phosphatase (AP) activity, calcium, phosphorus, triglycerides, and cholesterol were determined using the ADVIA® 1800 Clinical Chemistry analyzer (Siemens, Tarrytown, NY) utilizing specific reagents: ALPAMP, CA_2, IP, TRIG, and CHOL_2. These methods were available and applied to all samples collected at Sanford Research. Inorganic pyrophosphate (PP_i) was measured in lithium heparin plasma according to the method described previously [24 (link)]. Samples collected at the NYIT-COM were analyzed following the manual protocols in a plate reader (BioTek Instruments, Winooski, VT) operated in the kinetic (AP) or end-point mode (cholesterol, triglycerides, calcium, and phosphorus). PP_i concentration in those samples were determined based on luminescent detection of the ATP product of an ATP sulfurylase reaction, which uses PP_i as a substrate [25 (link)].

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Romanelli F., Corbo A., Salehi M., Yadav M.C., Salman S., Petrosian D., Rashidbaigi O.J., Chait J., Kuruvilla J., Plummer M., Radichev I., Margulies K.B., Gerdes A.M., Pinkerton A.B., Millán J.L., Savinov A.Y, & Savinova O.V. (2017). Overexpression of tissue-nonspecific alkaline phosphatase (TNAP) in endothelial cells accelerates coronary artery disease in a mouse model of familial hypercholesterolemia. PLoS ONE, 12(10), e0186426.

Publication 2017

ADVIA® 1800 Clinical Chemistry analyzer

Alkaline phosphatase Atp sulfurylase Blood Calcium Cholesterol Heparin Kinetic Lithium Luminescent Mice Phosphorus Plasma Pyrophosphate Right ventricle Triglycerides Venous blood

Corresponding Organization : University of Pennsylvania

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Alkaline phosphatase (AP) activity
Calcium
Phosphorus
Triglycerides
Cholesterol
Inorganic pyrophosphate (PPi)

control variables

Venous blood collected from the right ventricle of anesthetized mice
Mice were fasted at least 5 hours before blood collection

positive controls

None specified

negative controls

None specified

Annotations

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