Calcium assay kit

Manufactured by Nanjing Jiancheng

Sourced in China

The Calcium assay kit is a laboratory tool designed to quantitatively measure the calcium content in various samples. It provides a reliable and standardized method for calcium determination, allowing researchers to assess calcium levels accurately.

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Spelling variants of this product

Calcium colorimetric assay kit (4 citations)

27 protocols using calcium assay kit

Urine Analysis in Rat Metabolic Cages

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On Day 0 (for identifying the model establishment) or Day 28 (for evaluating the effects of Huashi Pill), the rats were fed in metabolic-cages and the 24-hour urine was harvested by incubating with 0.02% sodium azide (SinoPharm Co., Ltd., Shanghai, China) to prevent the appearance of bacteria. The urinary pH and urinary volume were evaluated and then the urine was used for the other urine biochemical analysis. The urine protein (urine protein quantitative kit, Cat. No. C035-2, Nanjing Jiancheng Bioengineering Inst., Nanjing, China), uric acid (uric acid assay kit, Cat. No. C012-1, Nanjing Jiancheng Bioengineering Inst.), calcium (calcium assay kit, Cat. No. C004-3, Nanjing Jiancheng Bioengineering Inst.), magnesium (magnesium assay kit, Cat. No. C005, Nanjing Jiancheng Bioengineering Inst.), phosphorus (phosphate assay kit, Cat. No. C006, Nanjing Jiancheng Bioengineering Inst.) were examined by utilizing the commercial kit on the semiautomatic photometer (Mode: 750, Hitachi, Tokyo, Japan) following to the instructions of the manufacturers.
Moreover, the levels of serum urea nitrogen (BUN) and creatinine (Cr) were also evaluated using the urea assay kit (Cat. No. C013-1, Nanjing Jiancheng Bioengineering Inst.) and creatinine assay kit (Cat. No. C011-2, Nanjing Jiancheng Bioengineering Inst.), according to the manufacturer’s instructions.

Yang A., Guo H., Fu M, & Liu M. (2019). Inhibitive Effects of Huashi Pill on Formation of Renal Stones by Modulating Urine Biochemical Indexes and Osteopontin in Renal Stone Rat Models. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 8335-8344.

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Biomimetic Mineralization of Microscaffolds

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To measure the weight increase ratio, the dry microscaffolds were weighted (W_d) and immersed into SBF for 7 day at 37 ℃. The obtained mineralized microscaffolds were weighted (W_m) again. The weight increase (%) was calculated by the equation:

Weight increase (%) = [(W_{m} - W_{d}) / W_{d}] \times 100 %

To analyze the calcium content in the microscaffolds after biomimetic mineralization, the mineralized microscaffolds were immersed in 0.5 mol/L acetic acid for 12 h. The calcium content was then calculated using a calcium assay kit (Jiancheng, Nanjing, China) according to the manufacturer’s instructions.

Yang Y., He H., Miao F., Yu M., Wu X., Liu Y., Fu J., Chen J., Ma L., Chen X., Peng X., You Z, & Zhou C. (2024). 3D-printed PCL framework assembling ECM-inspired multi-layer mineralized GO-Col-HAp microscaffold for in situ mandibular bone regeneration. Journal of Translational Medicine, 22, 224.

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Quantifying Plant Stress Signaling Molecules

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To test Ca²⁺, H₂O_2, ABA, PLA₂, NO, ABA and ET, adventitious roots (0.5 g, for NO and H₂O₂ assays, 0.1 g) were ground into homogenate on the rice with 4.5 mL PBS. The supernatant was collected at 4 °C for subsequent experiment after centrifugation at 3000 rpm for 20 min. Then the content of Ca²⁺, H₂O₂, NO was measured by Calcium Assay Kit, Hydrogen Peroxide assay kit (colorimetric method) and Nitric Oxide assay kit (Nitrate reductase method) (Nanjing Jiancheng Bioengineering Research Institute, Nanjing, China), and the content of ABA, PLA₂, ET were measured by Plant Abscisic Acid Elisa Kit, Plant PLA₂ Elisa Kit and Plant ET Elisa Kit (Senbejia Nanjing Biotechnology Co., Ltd, Nanjing, China) according the manufactures’ directions.

Wang S.H., Liang W.X., Lu J., Yao L., Wang J, & Gao W.Y. (2020). Penicillium sp. YJM-2013 induces ginsenosides biosynthesis in Panax ginseng adventitious roots by inducing plant resistance responses. Chinese Herbal Medicines, 12(3), 257-264.

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Mandibular Microstructure in p75NTR Mice

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All mandibles of the 1‐month‐old (n = 5) and 4‐month‐old (n = 5) male wild‐type (WT) mice and their male p75NTR^−/− littermates were dissected free of soft tissue and scanned by micro‐CT (viva CT 40, Scanco Medical AG; Switzerland). Three‐dimensional (3D) microstructural parameters were calculated using MicroView software version 2.2. Alveolar bone microstructural parameters were measured directly from the region of interest (ROI) which was manually established in the inter‐radicular septa of the mandibular first molar (M1). The sampling region started coronally from the M1 inter‐radicular septa to root apex direction after 50 continuous scan slices, and 50 continuous scan slices with the same shape and tissue volume were considered as the ROI in each specimen. Three‐dimensional micro‐architecture parameters of the ROI were also evaluated.
Serum calcium and phosphorus were measured with a Calcium Assay Kit (Nanjing Jiancheng Bioengineering Institute, China) and a Phosphate Assay Kit (Nanjing Jiancheng Bioengineering Institute, China), respectively. Serum parathyroid hormone (PTH) and calcitonin (CT) were detected using a PTH ELISA Kit (RayBiotech, USA) and a Calcitonin Assay Kit (Cloud‐Clone Corp. China), respectively.

Wang Y., Yang K., Li G., Liu R., Liu J., Li J., Tang M., Zhao M., Song J, & Wen X. (2020). p75NTR−/− mice exhibit an alveolar bone loss phenotype and inhibited PI3K/Akt/β‐catenin pathway. Cell Proliferation, 53(4), e12800.

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Quantifying Mineralized Nodules and Calcium Content

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Von Kossa: Von Kossa assay was performed to determine the mineralized nodules on microtissues as described 29 (link). After washing twice with PBS, samples were fixed in 4% paraformaldehyde for 1 h. This was followed by rinsing with double distilled water and incubating with 2% AgNO₃ in darkness for 10 min, followed by exposure to UV light for 15 min. Finally, samples were observed under a camera equipped with a close-up filter.
Calcium content: Thirty microtissues was harvested at the corresponding time points (12 h, day 7, day 14, day 21) after changing the osteogenic medium. The samples were washed twice with PBS and immersed in 0.5 mL of 10% acetic acid to isolate calcium from the microtissues. The samples were then tested using the Calcium Assay Kit (Jiancheng, Nanjing, China). Finally, specimens were observed under a DU 730 UV/Vis spectrophotometer (Beckman Coulter) and the absorbance at 610 nm was measured (optical density, OD).

Luo C., Fang H., Zhou M., Li J., Zhang X., Liu S., Zhou C., Hou J., He H., Sun J, & Wang Z. (2019). Biomimetic open porous structured core-shell microtissue with enhanced mechanical properties for bottom-up bone tissue engineering. Theranostics, 9(16), 4663-4677.

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Quantification of Cellular Calcium Levels

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Calcium concentrations of the cells and rat aortic rings were quantified using a commercially available Calcium Assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), by following previously established protocols.^{19 (link),20 (link)} Briefly, after the cells and aortic rings were harvested and washed with phosphate-buffered saline, they were treated with 0.6 N HCl overnight at 4°C. Calcium concentrations in the supernatant were determined by the o-cresolphthalein complexone method. A Bicinchoninic Acid Protein Assay kit (Aspen Biotechnology Co., Ltd., Wuhan, China) was used to evaluate protein concentrations to normalize calcium concentrations.^{19 (link),20 (link)}

Zang H., Liu Y., Teng Q., Hua J., Peng D, & Wang P. (2024). Phosphonoformic acid reduces hyperphosphatemia-induced vascular calcification via Pit-1. The Journal of International Medical Research, 52(1), 03000605231222156.

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MSC Differentiation with UHMWPE and USC-EVs

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BMSCs were isolated from the femurs and tibias of 4-week-old male C57BL/6 mice. BMSCs were seeded into 48-well plates, and 24 h later, the old culture medium was changed to osteogenesis induction medium (Cyagen Biosciences Inc., Guangzhou, China) containing UHMWPE particles (1.0 mg/mL) with or without USC-EVs (100 μg/mL). Four days later, conditioned medium (CM) was obtained and assayed with an alkaline phosphatase (ALP) assay kit and calcium assay kit according to the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). After 12 days of induction, the cells were fixed and stained with an Alizarin Red S (ARS) solution (Cyagen Biosciences Inc., Guangzhou, China). Images were captured under an inverted microscope (Leica DMI6000B, Solms, Germany).

Li H., Fan X.L., Wang Y.N., Lu W., Wang H., Liao R., Zeng M., Yang J.X., Hu Y, & Xie J. (2021). Extracellular Vesicles from Human Urine-Derived Stem Cells Ameliorate Particulate Polyethylene-Induced Osteolysis. International Journal of Nanomedicine, 16, 7479-7494.

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Nephrolithiasis Induction and Analysis

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After MS modeling was successfully established at 12 weeks, drug administration was carried out to induce nephrolithiasis in rats. The urine samples of rats were collected in metabolic cages to perform a routine urine test (URITEST-500B urine analyzer, Guangxi, China) and measure urinary oxalate (Oxalate Assay kit, Nanjing Jiancheng Technology, China) and urinary calcium (calcium assay kit, Nanjing Jiancheng Technology, China) levels once a week.

He Q., Tang Y., Li Y., Wang F., Bao J, & Gupta S. (2022). A pilot dynamic analysis of formative factors of nephrolithiasis related to metabolic syndrome: evidence in a rat model. Renal Failure, 44(1), 1134-1143.

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Measuring Calcium Content in HVSMCs

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Calcium assay kit (C004-2-1, Jiancheng, China) was used to examine the Ca²⁺ content in HVSMCs. HVSMCs with different intervention were collected and subsequently lysed and centrifuged (4℃, 12,000×g, 5 min). The supernatant was collected to measure Ca²⁺ content according to the instructions [17 (link)].

Li H., Zhang C, & Liu Q. (2023). Lumican silencing ameliorates β-glycerophosphate-mediated vascular smooth muscle cell calcification by attenuating the inhibition of APOB on KIF2C activity. Open Medicine, 18(1), 20230790.

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Characterization of Extracellular Matrix

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After 5 days of mechanical stimulus, the MC3T3-E1 cells were eliminated according to our established method [36 (link),37 (link)] (showed in Additional file 1), then the ECMs coated on the loading dishes were prepared (Additional file 1: Figure S1).

Measure hydroxyproline content. The ECMs on the loading dishes were hydrolyzed with hydrolysis buffer and the hydroxyproline content were detected with the Chloramine-T Hydroxyproline Assay Kit contain lysis buffer (Nanjing Jiancheng Biotechnology Institute Co., Ltd., Nanjing, China.) according to the manufacturer’s protocol.

Glycosaminoglycan (GAG) analysis. The ECMs were lysed with lysis buffer and GAG levels were assayed with Dimethyl-methylene Blue GAG Assay Kit contain lysis buffer (Xianmen Maiwei Biotechnology Co., Ltd., Xiamen, China.). The absorbance or optical density (OD) at 660 nm was regarded as relative level of GAG in the ECMs. Results were expressed as relative to control group (the ECM of unstrained cells on loading dishes).

Calcium deposition assay. After the ECMs were treated overnight with 0.1 M HCl, the ECM-deposited calcium content of the dishes was measured with the Calcium Assay Kit (Nanjing Jiancheng Biotechnology Co., Inc.) using the methylthymol blue complexon method according to the manufacturer’s instructions.

Zeng Q., Guo Y., Liu Y., Li R., Zhang X., Liu L., Wang Y., Zhang X, & Zou X. (2015). Integrin-β1, not integrin-β5, mediates osteoblastic differentiation and ECM formation promoted by mechanical tensile strain. Biological Research, 48(1), 25.

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