Alizarin Red S was performed to visualize the formation of mineralized matrix (Shanghai Gefan Biological Technology Co., Ltd, Shanghai, China). The calcium content was determined using a calcium colorimetric assay kit (Nanjing Jiancheng Biological Engineering Institute, Nanjing, China). Alkaline phosphatase (ALP) activity was measured using an ALP kit (Beyotime Institute of Biotechnology, Shanghai, China). Caspase-3 activity was analyzed using a caspase-3 assay kit (Beyotime Institute of Biotechnology, Shanghai, China).
Calcium colorimetric assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The Calcium colorimetric assay kit is a laboratory equipment product designed to measure the concentration of calcium in various samples. The kit utilizes a colorimetric method to quantify the calcium content, providing accurate and reliable results for research and analytical applications.
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Lab products found in correlation
4 protocols using calcium colorimetric assay kit
1
Mineralization and Cell Viability Assays
2
Extracellular Matrix Mineralization in BMSCs
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12 days after culturing under NP, the extracellular matrix mineralization of BMSCs were evaluated by the alizarin red S staining and calcium contents. The BMSCs were fixed in 4% paraformaldehyde for 15 min and gently rinsed with PBS. Finally, the sample were then incubated with 40 mM alizarin red S (ARS) staining solution (Sigma) for 30 min at room temperature. Images were recorded with an Olympus MVX10 MacroView (Japan). For quantitative analysis, calcium contents were measured using a calcium colorimetric assay kit (Nanjing Jiancheng, China) according to the manufacturer’s instructions. All samples were examined at less three times.
3
Quantification of H2O2, NO, and Calcium
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A Quantitative Assay Kit (Nanjing Jiancheng, Beijing, China) was used for the determination of H2O2 and nitric oxide (NO) in the control and GhRbohD-silenced cotton plants, referring to the manual for detailed operation steps.
The calcium content of the control and GhRbohD-silenced plants’ leaves were determined with a Calcium Colorimetric Assay Kit (Nanjing Jiancheng, Beijing, China). The samples’ absorbance was measured at 575 nm, and the concentrations of calcium were calculated based on the formula: C = Sa/Sv (Sa: amount of calcium in unknown sample (µg) from standard curve; Sv: sample volume (µL) added into the wells; C: concentration of calcium in sample). Three independent biological and technical repeats were performed.
The calcium content of the control and GhRbohD-silenced plants’ leaves were determined with a Calcium Colorimetric Assay Kit (Nanjing Jiancheng, Beijing, China). The samples’ absorbance was measured at 575 nm, and the concentrations of calcium were calculated based on the formula: C = Sa/Sv (Sa: amount of calcium in unknown sample (µg) from standard curve; Sv: sample volume (µL) added into the wells; C: concentration of calcium in sample). Three independent biological and technical repeats were performed.
4
Reproductive Hormone Assay Protocol
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For intervention, oestradiol benzoate for injection (Ningbo Sansheng Pharmaceutical Co., Ltd., batch number: 120904), oxytocin for injection (Shanghai Hefeng Factory, batch number: 130115), and Yuanhu Zhitong tablets (Tianjin Tongrentang Group Limited by Share Ltd., batch number: c91017) were used.
For outcome, prostaglandin E2 assay kit, prostaglandin F2α assay kit, nitric oxide assay kit (nitrate reductase method), and calcium colorimetric assay kit (Nanjing Jiancheng Bioengineering Institute); YXJ-type high speed centrifuge (Changzhou Jiancheng Teaching Instrument Factory); OLYMPUS U-CMAD3 optical microscope; Japanese OLYMPUS, OLYMPUS C5060-ADU optical camera; Japanese OLYMPUS; gas bath thermostats oscillator (Jintan Ronghua Instrument Manufacturing Co., Ltd.); and Infinite 200 PRO automatic enzyme mark instrument (Tecan Group, Ltd.) were applied.
For outcome, prostaglandin E2 assay kit, prostaglandin F2α assay kit, nitric oxide assay kit (nitrate reductase method), and calcium colorimetric assay kit (Nanjing Jiancheng Bioengineering Institute); YXJ-type high speed centrifuge (Changzhou Jiancheng Teaching Instrument Factory); OLYMPUS U-CMAD3 optical microscope; Japanese OLYMPUS, OLYMPUS C5060-ADU optical camera; Japanese OLYMPUS; gas bath thermostats oscillator (Jintan Ronghua Instrument Manufacturing Co., Ltd.); and Infinite 200 PRO automatic enzyme mark instrument (Tecan Group, Ltd.) were applied.
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