To obtain the cell supernatant, collect 500 μl of the cell culture medium after intervening with cardiac fibroblasts, and centrifuge it at 4 °C and 6000 rpm for 10 min to remove any cellular debris. The concentration of the Col1 protein in cell supernatant was determined using the I-type collagen α1 enzyme-linked immunosorbent assay kit, following the instructions provided by the manufacturer (E-EL-R0041c, Elabscience).
E el r0041c
Manufactured by Elabscience
The E-EL-R0041c is a laboratory equipment product manufactured by Elabscience. It is a device used for the detection and analysis of proteins. The core function of this product is to facilitate protein detection and quantification in research and laboratory settings.
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Lab products found in correlation
2 protocols using e el r0041c
1
Quantifying Collagen-1 in Cardiac Fibroblast Supernatant
2
Quantification of Lung Collagen and Hydroxyproline
Check if the same lab product or an alternative is used in the 5 most similar protocols
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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).
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