Heart samples were weighed, homogenized in deionized water, and hydrolyzed overnight in 6N HCl at 120°C. Lysates were transferred to and desiccated in 96-well plates and reconstituted in deionized water. After incubation with 50 mM chloramine T (Sigma-Aldrich) followed by 1 M dimethylaminobenzaldehyde (Sigma-Aldrich), the OD values were read at 570 nm. The concentration of hydroxyproline was determined by a 1–100 µg/ml standard curve of hydroxyproline (Sigma-Aldrich) and normalized to starting heart tissue mass (Wu et al., 2014 (link)).
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Quantifying Collagen via Hydroxyproline Assay
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Corresponding Organization : Johns Hopkins University
Other organizations : Johns Hopkins Medicine
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Variable analysis
independent variables
- Heart samples weighed
- Heart samples homogenized in deionized water
- Heart samples hydrolyzed overnight in 6N HCl at 120°C
- Lysates incubated with 50 mM chloramine T (Sigma-Aldrich)
- Lysates incubated with 1 M dimethylaminobenzaldehyde (Sigma-Aldrich)
- OD values read at 570 nm
- Concentration of hydroxyproline determined by a 1–100 µg/ml standard curve of hydroxyproline (Sigma-Aldrich)
- Hydroxyproline concentration normalized to starting heart tissue mass
- Deionized water used for homogenization and reconstitution
- 1–100 µg/ml standard curve of hydroxyproline (Sigma-Aldrich)
- Not explicitly mentioned
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