HYP is one of the main components of collagen in the body and is distributed mostly in skin, tendons, cartilage, and blood vessels. Thus, HYP content is an important indicator reflecting the metabolism and fibrosis degree of collagen tissue. The HYP content in mouse lung tissue was detected with an HYP assay kit (Solarbio, Beijing).
Hyp assay kit
Manufactured by Solarbio
Sourced in China
The HYP assay kit is a laboratory product used to detect and quantify the levels of the chemical compound hydroxyproline (HYP) in various biological samples. The kit provides the necessary reagents and protocols to perform this specific analysis.
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Lab products found in correlation
Synergy lx microplate reader, by Agilent Technologies (1 mentions)
E el r0041c, by Elabscience (1 mentions)
Triton x 100, by Sinopharm (1 mentions)
Tryptone, by Thermo Fisher Scientific (1 mentions)
Soya peptone, by Sinopharm (1 mentions)
Ch3 ch2 11so4na, by Sinopharm (1 mentions)
Cell counting kit 8 cck 8, by Apexbio (1 mentions)
8 protocols using hyp assay kit
1
Quantifying Collagen Metabolism in Mouse Lungs
2
Quantification of Lung Collagen and Hydroxyproline
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The expression of type I and III collagen major isoforms (Col1a1 and Col3a1) was detected by ELISA kits (E-EL-R0041c and E-EL-R0235c, Elabscience, Wuhan, CN). Five rats for each test group were used. In brief, 100 mg of frozen lung tissue of each rat was ground in liquid nitrogen and resuspended in 1mL PBS. After repeated freeze-thaw, homogenate supernatant was obtained by centrifugation at 5000 g for 5 minutes at 4°C. After that, standard ELISA procedure was carried out according to the manufacturer’s instruction and absorbance at 450 nm was measured by a Synergy LX microplate reader (BioTek, USA).
The hydroxyproline content in lung tissue homogenate was detected by a HYP assay kit (BC0255, Solarbio, Beijing, CN). Five rats for each test group were used. In brief, 200 mg of frozen lung tissue of each rat was cut into very small pieces and digested 4–6 hours at 110°C within 2 mL of digestion buffer. After adjusting pH to 6–8 and making total volume to 4 mL with sterile water, the supernatant was obtained by centrifugation at 16,000 rpm for 20 minutes at 25°C. After that, HYP concentration in the homogenate was determined according to the manufacturer’s instruction and absorbance at 560 nm was measured by a Synergy LX microplate reader (BioTek, USA).
The hydroxyproline content in lung tissue homogenate was detected by a HYP assay kit (BC0255, Solarbio, Beijing, CN). Five rats for each test group were used. In brief, 200 mg of frozen lung tissue of each rat was cut into very small pieces and digested 4–6 hours at 110°C within 2 mL of digestion buffer. After adjusting pH to 6–8 and making total volume to 4 mL with sterile water, the supernatant was obtained by centrifugation at 16,000 rpm for 20 minutes at 25°C. After that, HYP concentration in the homogenate was determined according to the manufacturer’s instruction and absorbance at 560 nm was measured by a Synergy LX microplate reader (BioTek, USA).
3
Hydroxyproline Quantification in Liver
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Hydroxyproline levels were detected using a HYP assay kit (Solarbio, Beijing, China; cat: BC0255). Liver tissues were cut into pieces and digested in the extraction solution at approximately 100 °C for 2–6 h, until the tissues were totally hydrolyzed. The PH of the mixture was adjusted to 10 using 10 M NaOH. The adjusted mixture was then centrifuged at 16,000 rpm for 30 min, and the supernatant was collected for testing. The OD value of each well was measured at 560 nm with a SpectraMax ABS PLUS reader. The HYP content was calculated as previously described [94 (link)].
4
Collagen Content Analysis in Lung
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The content of collagen in the lung homogenate was analyzed using a hydroxyproline (HYP) assay kit (Solarbio, China).
5
Hydroxyproline Assay in Lung
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The HYP content of the lung homogenate was detected using an HYP assay kit (Solarbio, China) according to the manufacturer’s instruction. The absorbance of the samples was measured at 550 nm.
6
Quantifying Hydroxyproline in Tissue
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An HYP assay kit (BC0255, Solarbio, China) was used to test the variation in the HYP content in the tissue samples. First, the tissue was accurately weighed. After hydrolysis, the pH was adjusted to neutral, and the supernatant was centrifuged with activated carbon. The reagent was added and then incubated in water at 60°C for 15 min. After 10 min at 3,500 rpm, the supernatant was taken for spectrophotometry at 550 nm.
7
Glycosaminoglycan ELISA and Cell Assays
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Vancomycin was purchased from Aladdin (Shanghai, China), Triton X100 and CH3(CH2)11SO4Na from Sinopharm Chemical Reagent Co., Ltd. (China), Pig glycosaminoglycan (GAG) ELISA Kit from Jianglai Biotechnology Co., Ltd (Shanghai China), Hyp assay kit from Beijing solarbio science & technology Co., Ltd. (China). Agar from Sangon Biotech Co., Ltd. (Shanghai, China), TRYPTONE from OXOID (Shanghai, China), and soya peptone from Sinopharm Chemical Reagent Co., Ltd. (China). DNA kit from CW Biotech (Beijing, China), Live/dead cell staining dye was purchased from BioVision (USA), and Cell Counting Kit-8 (CCK8) from APExBIO Technology LLC (USA).
8
Collagen Metabolism Evaluation in Mouse Lung
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HYP is one of the main components of collagen in the body and is distributed mostly in skin, tendons, cartilage, and blood vessels. Thus, HYP content is an important indicator reflecting the metabolism and fibrosis degree of collagen tissue. The HYP content in mouse lung tissue was detected with an HYP assay kit (Solarbio, Beijing).
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