May 16, 2019
Yale - Beta hydroxybutyrate (Cobas)
- Gary Cline1,
- John Stack1
- 1Yale University
- Mouse Metabolic Phenotyping CentersTech. support email: [email protected]
Protocol Citation: Gary Cline, John Stack 2019. Yale - Beta hydroxybutyrate (Cobas). protocols.io https://dx.doi.org/10.17504/protocols.io.y2bfyan
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 11, 2019
Last Modified: May 16, 2019
Protocol Integer ID: 21283
Keywords: Beta hydroxybutyrate, beta hydroxybutyrate, coba, serum
Abstract
Summary:
Procedure used to determine the concentration of β-Hydroxybutyrate in blood, serum, and plasma. β-Hydroxybutyrate is measured by the oxidation to acetoacetate, followed by reduction of an indicator dye (monitored at 505 nm) by NADH.
Materials
MATERIALS
ß-Hydroxybutyrate ControlsPointe Scientific Inc.Catalog #H7587-CTL
ß-Hydroxybutyrate Reagent (test kit includes standard). Pointe Scientific Inc.Catalog #H7587-58
Reagent Preparation:
ß-Hydroxybutyrate Controls: As supplied by vendor
ß-Hydroxybutyrate Reagent and Standard: As supplied by vendor
Troubleshooting
Before start
Analysis by automated system Cobas Mira Plus.
Calibrate Cobas for β-Hydroxybutyrate by running a β-Hydroxybutyrate standard and three ß-Hydroxybutyrate controls.
Sample handling as performed by the Cobas Mira Plus.
a) Pipette 3 µL of sample into cuvette.
b) Add 105µL of ß-Hydroxybutyrate Reagent.
c) Mixture is incubated at 37˚C for 10 minutes.
d) Absorbance is measured at 505 nm.