We do not have data supporting frozen samples and recommend using fresh samples. However, provided this is not available, it would be prudent to compare the readings from a sample frozen versus fresh to see if there is any impact on the determination.
MAK357
Hydroxyproline Assay Kit (Perchlorate-Free)
sufficient for 100 colorimetric tests
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Recommended Products
usage
sufficient for 100 colorimetric tests
detection method
colorimetric
relevant disease(s)
rheumatological diseases; cancer; neurological disorders; cardiovascular diseases
storage temp.
−20°C
General description
The new Hydroxyproline Assay Kit, MAK463, is now available! Hydroxyproline ((2S,4R)-4-hydroxyproline, Hyp) is a nonproteinogenic amino acid formed by the enzyme prolyl-4-hydroxylase, which catalizes the post-translational modification of polypeptide proline residues following protein synthesis. In animals, hydroxyproline is found almost entirely in collagen, which contains approximately 12-14% hydroxyproline by mass (the fibrilous protein elastin also contains small quantities of hydroxyproline). Hydroxyproline in tissue hydrolysates is thus a direct measure of the amount of collagen present. Collagen is the most abundant protein in mammals and many diseases are believed to affect collagen turnover, including tumor invasion and metastasis, rheumatoid arthritis, cardiopulmonary fibrosis and muscular dystrophy.
Features and Benefits
Perchlorate (PCA)-Free
Suitability
Suitable for the estimation of hydroxyproline concentration in tissue homogenates (i.e. muscle, liver, lung, etc.) and human or animal biological fluids (serum, urine).
Principle
The classical assay for determination of Hyp is based upon oxidation of Hyp to a pyrrole intermediate, followed by reaction with Ehrlich′s reagent dissolved in concentrated perchloric acid (HClO4). Perchloric acid is a hazardous material that is both toxic and highly reactive, requiring special handling and waste-disposal protocols. The Hydroxyproline Assay Kit (Perchlorate-Free) uses a proprietary acidic developer solution to accurately measure Hyp in tissue and protein/peptidase hydrolysates without the use of hazardous perchlorates. It is a quick and convenient protocol, yielding a brightly-colored chromophore with an absorbance peak at 560 nm. The assay can detect a minimum of 0.05 ug hydroxyproline per well in a 96-well format and generates colorimetric results that are virtually identical to the classical perchloric acid-based method.
replaced by
Product No.
Description
Pricing
signalword
Danger
Hazard Classifications
Aquatic Chronic 3 - Eye Dam. 1 - Met. Corr. 1 - Resp. Sens. 1 - Skin Corr. 1A - Skin Sens. 1 - STOT SE 3
target_organs
Central nervous system
Storage Class
8A - Combustible corrosive hazardous materials
flash_point_f
188.6 °F
flash_point_c
87 °C
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Hi, I was wondering if the tissue has to be fresh before homogenization and if freeze and thaw would affect the accuracy of the assay
1 answer-
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Can Product MAK357, Hydroxyproline Assay Kit Perchlorate-Free, be used to measure the hydroxyproline levels in cell culture supernatants?
1 answer-
The kit is optimized for use in determination of hydroxyproline in biological samples (e.g. plasma, serum, cell lysate, tissue samples.) This kit has not been tested for use with cell culture supernatant. The presence of phenol red may interfere with results. In addition, the cultured cell line may not secrete sufficient hydroxyproline to be detected. It would be up to the end user to determine suitability.
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