The concentration of hepatic hydroxyproline from each mouse was determined with a commercial hydroxyproline testing kit (Jiancheng, Nanjing, China) according to the manufacturer’s instruction. In brief, frozen liver tissue (approximate 30 mg) was heated in 6 mM HCl at 120 °C for 4 h in sealed tubes. The samples were then centrifuged, and 10 ml of supernatant was mixed with 100 ml of chloramine-T solution (1.4% chloramine-T, 10% N-propanol, and 80% citrate-acetate buffer). This mixture was incubated for 20 min at room temperature. After that, Ehrlich’s solution was added, and the samples were incubated at 65 °C for 20 min. The absorbance was measured at 550 nm, and the concentration was determined by the standard curve created by cis-4-hydroxy-L-proline (Sigma-Aldrich, USA).
Cis 4 hydroxy l proline
Manufactured by Merck Group
Sourced in United States
Cis-4-hydroxy-L-proline is a chemical compound that is commonly used in various laboratory procedures. It is a non-proteinogenic amino acid that can be utilized in the synthesis of peptides and other organic compounds. The core function of this product is to serve as a building block in chemical reactions and analyses, though its specific applications may vary depending on the research or industrial context.
Automatically generated - may contain errors
Lab products found in correlation
Dna oligonucleotide, by Integrated DNA Technologies (1 mentions)
Yeast trnaphe, by Merck Group (1 mentions)
Spermine s4264, by Merck Group (1 mentions)
L azetidine 2 carboxylic acid, by Merck Group (1 mentions)
L proline, by Merck Group (1 mentions)
D proline, by Merck Group (1 mentions)
Trans 4 hydroxy l proline, by Merck Group (1 mentions)
Chloramine t, by Merck Group (1 mentions)
4 protocols using cis 4 hydroxy l proline
1
Hepatic Hydroxyproline Quantification
2
Proline Analogs and Polyamine Purification
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Proline and its analogs, α-methyl-l -proline, 3,4-dehydro-dl -proline, cis-4-hydroxy-l -proline and l -azetidine-2-carboxylic acid, as well as spermidine (catalog number S2626), spermine (S4264) and putrescine (P5780), and yeast tRNAPhe were purchased from Sigma. All DNA oligonucleotides were purchased from either Integrated DNA Technologies or Eurofins MWG Operon. The 3′-biotin-labeled RNA oligonucleotide used for affinity purification of yeast tRNAPro was also purchased from Eurofins MWG Operon; yeast tRNALys was obtained from tRNA Probes (College Station, TX, USA).
3
Proline Stereoisomers Characterization
4
Quantifying Hydroxyproline in Cells
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Hydroxyproline measurement was done using a modified protocol that was described elsewhere [5] . Briefly, 20 mg of frozen fat or 3T3-L1 cells were heated in 6 N HCl at 110 °C overnight in sealed tubes. The samples were then heated at 110 °C until dried. Each sample was incubated with chloramine-T (Sigma Aldrich) at room temperature for exactly 20 min and then with p-dimethylaminobenzaldehyde (Fisher Scientific) at 60 °C for 15 min. The absorbance was read at 540 nm and the concentration was determined by the standard curve created with cis-4-hydroxy-L-proline (Sigma-Aldrich).
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