Quantichrome calcium assay kit

Manufactured by Gentaur

Sourced in Belgium

The QuantiChrome Calcium Assay Kit is a laboratory tool designed to quantify the concentration of calcium in various samples. It utilizes a colorimetric method to measure the calcium content, providing a reliable and accurate means of analysis.

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Alizarin red s, by Merck Group (2 mentions) S8045, by Merck Group (2 mentions) Sodium dodecyl sulfate (sds), by Merck Group (2 mentions) Dm il led microscope, by Leica (2 mentions) Alizarin red s, by Leica (1 mentions)

9 protocols using quantichrome calcium assay kit

Quantification of Osteoblast Mineralization

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Cells grown on 48‐well plates were washed twice with PBS and decalcified with 0.6 mol/l HCl for 24 hrs at 37°C. Calcium content of the supernatants was determined by the QuantiChrome Calcium Assay Kit (Gentaur, Brussels, Belgium). After decalcification, cells were solubilized with a solution of NaOH 0.1 mol/l and SDS 0.1%, and protein content of samples was measured with (bicinchoninic acid) BCA protein assay kit (Pierce, Rockford, IL, USA). Calcium content of the cells was normalized to protein content and expressed as μg/mg protein. Mineralization was also determined by Alizarin Red staining. Briefly, cells grown on 48‐well cell culture plates were washed with PBS and fixed with 4% paraformaldehyde. Plates were incubated for 10 min. in RT. Cells were washed with PBS and then stained with 2% Alizarin Red solution for 2 min. After the incubation cells were washed three times with distilled water and photos were taken under light microscope.

Becs G., Zarjou A., Agarwal A., Kovács K.É., Becs Á., Nyitrai M., Balogh E., Bányai E., Eaton J.W., Arosio P., Poli M., Jeney V., Balla J, & Balla G. (2015). Pharmacological induction of ferritin prevents osteoblastic transformation of smooth muscle cells. Journal of Cellular and Molecular Medicine, 20(2), 217-230.

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Calcium-Induced Vascular Calcification Assay

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At approximately 80% confluence, VICs were cultured in a calcification medium with or without phenol red supplemented with 2.5 mmol/L inorganic phosphate and 1.8 mmol/L calcium chloride for 5 days. For time-dependent experiments, the first day of culturing in calcification medium was considered as day 0.
VICs were washed twice with phosphate-buffered saline (PBS) and decalcified using 0.6 mol/L hydrochloric acid (HCl). The calcium contents of the supernatants were assigned by QuantiChrome Calcium Assay Kit (Gentaur; 65-DICA-500), normalized to protein content.
Alizarin Red S (Sigma Aldrich; A5533) stainings were used to visualize calcium deposition. Cells were fixed with 3.7% formaldehyde for 10 min at 4 °C and stained with 2% Alizarin Red S. Stained cells were visualized using Leica DMIL LED microscope, Leica DMC4500 camera with Leica application suite LAS Software 4.9.0× and 10× magnification). Calcified regions were evaluated by ImageJ software.

Combi Z., Potor L., Nagy P., Sikura K.É., Ditrói T., Jurányi E.P., Galambos K., Szerafin T., Gergely P., Whiteman M., Torregrossa R., Ding Y., Beke L., Hendrik Z., Méhes G., Balla G, & Balla J. (2023). Hydrogen sulfide as an anti-calcification stratagem in human aortic valve: Altered biogenesis and mitochondrial metabolism of H2S lead to H2S deficiency in calcific aortic valve disease. Redox Biology, 60, 102629.

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Quantifying Calcification in Valvular Cells

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Valvular interstitial cells grown on cell culture plates until 90% confluent, they were washed twice with PBS and decalcified using 0.6 mol/L HCl for 1 hour. Calcium content of the supernatants was determined by QuantiChrome Calcium Assay Kit (Gentaur). After decalcification, cells were solubilized with a solution of NaOH 0.1 mol/L and SDS 0.1%. Calcium content of the cells was normalized to protein content and expressed as μg/mg protein. Calcium was determined by QuantiChrome Calcium Assay Kit (BioAssays System; DICA-500) and normalized to protein content. Alizarin Red S (Sigma Aldrich; A5533) staining was employed to visualize the calcium deposition. Plates were fixed with 3 % paraformaldehyde, and stored at 4°C for 10 minutes, and stained with a 2 % solution of Alizarin Red S. All calcific nodules in each well were then manually counted under a microscope.

Sikura K.É., Potor L., Szerafin T., Zarjou A., Agarwal A., Arosio P., Poli M., Hendrik Z., Méhes G., Oros M., Posta N., Beke L., Fürtös I., Balla G, & Balla J. (2019). POTENTIAL ROLE OF H-FERRITIN IN MITIGATING VALVULAR MINERALIZATION. Arteriosclerosis, thrombosis, and vascular biology, 39(3), 413-431.

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Quantifying Cellular Calcium Content

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Cells grown on 96-well plates were washed twice with DPBS and decalcified with HCl (30721, Sigma, 0.6 mol/L) for 30 min at room temperature. The Ca content of the HCl supernatants was determined by using a QuantiChrome Calcium Assay Kit (DICA-500, Gentaur, Kampenhout, Belgium). Following decalcification, cells were washed twice with DPBS and solubilized with a solution of NaOH (S8045, Sigma, 0.1 mol/L) and sodium dodecyl sulfate (11667289001, Sigma, 0.1%), and protein content of samples were measured using the BCA protein assay kit (23225, Pierce Biotechnology, Rockford, IL, United States). The Ca content of the cells was normalized to protein content and expressed as mg/mg protein. The observer who performed all the Ca measurements was blinded to the group assignment.

Tóth A., Csiki D.M., Nagy B J.r., Balogh E., Lente G., Ababneh H., Szöőr Á, & Jeney V. (2022). Daprodustat Accelerates High Phosphate-Induced Calcification Through the Activation of HIF-1 Signaling. Frontiers in Pharmacology, 13, 798053.

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Quantifying Cellular Calcium Content

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Cells grown on 96-well plates were washed twice with DPBS, and decalcified with HCl (30721, Sigma, St. Louis, MO, USA, 0.6 mol/L) for 30 min at room temperature. The Ca content of the HCl supernatants was determined using the QuantiChrome Calcium Assay Kit (DICA-500, Gentaur, Kampenhout, Belgium). Following decalcification, cells were washed twice with DPBS, and solubilized with a solution of NaOH (S8045, Sigma, St. Louis, MO, USA, 0.1 mol/L) and sodium dodecyl sulfate (11667289001, Sigma, St. Louis, MO, USA 0.1%), and the protein content of the samples was measured with the BCA protein assay kit (23225, Pierce Biotechnology, Rockford, IL, USA). The Ca content of the cells were normalized to the protein content, and expressed as mg/mg protein.

Balogh E., Chowdhury A., Ababneh H., Csiki D.M., Tóth A, & Jeney V. (2021). Heme-Mediated Activation of the Nrf2/HO-1 Axis Attenuates Calcification of Valve Interstitial Cells. Biomedicines, 9(4), 427.

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Quantifying Cellular Calcium Content

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Cells grown on 96-well plates were washed twice with DPBS, and decalcified with HCl (30721, Sigma, 0.6 mol/L) for 30 min at room temperature. Calcium content of the HCl supernatants was determined by QuantiChrome Calcium Assay Kit (DICA-500, Gentaur, Kampenhout, Belgium). Following decalcification, cells were washed twice with DPBS, and solubilized with a solution of NaOH (S8045, Sigma, Burlington, MA, United States, 0.1 mol/L) and sodium dodecyl sulfate (11667289001, Sigma, Burlington, MA, United States, 0.1%), and protein content of samples were measured with BCA protein assay kit (23225, Pierce Biotechnology, Rockford, IL, United States). Calcium content of the cells was normalized to protein content and expressed as μg/mg protein.

Szabó L., Balogh N., Tóth A., Angyal Á., Gönczi M., Csiki D.M., Tóth C., Balatoni I., Jeney V., Csernoch L, & Dienes B. (2022). The mechanosensitive Piezo1 channels contribute to the arterial medial calcification. Frontiers in Physiology, 13, 1037230.

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Alizarin Red S Staining for Calcification

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The VIC were cultured in a calcification medium (2.5‐mmol·L⁻¹ inorganic phosphate and 1.8‐mmol·L⁻¹ calcium–chloride) with or without phenol red for 5 days. Calcium content of the supernatants was determined by QuantiChrome Calcium Assay Kit (Gentaur), normalized to protein content, and expressed as μg·mg⁻¹ protein. Alizarin Red S staining was used to visualize the calcium deposition. Plates were fixed with 3.7% formaldehyde for 10 min followed by staining with a 2% solution of Alizarin Red S. Pictures from the stained cells were taken with a light microscope (×10 magnification; Leica DMIL LED microscope).

Sikura K.É., Potor L., Szerafin T., Oros M., Nagy P., Méhes G., Hendrik Z., Zarjou A., Agarwal A., Posta N., Torregrossa R., Whiteman M., Fürtös I., Balla G, & Balla J. (2019). Hydrogen sulfide inhibits calcification of heart valves; implications for calcific aortic valve disease. British Journal of Pharmacology, 177(4), 793-809.

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Quantification of Cellular Calcium Content

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Cells grown on 96-well plates were washed twice with DPBS and decalcified with HCl (30721, Sigma, 0.6 mol/L) for 30 min at room temperature. Ca content of the HCl supernatants was determined by QuantiChrome Calcium Assay Kit (DICA-500, Gentaur, Kampenhout, Belgium). Following decalcification, cells were washed twice with DPBS and solubilized with a solution of NaOH (S8045, Sigma, 0.1 mol/L) and sodium dodecyl sulfate (11667289001, Sigma, 0.1%), and the protein content of samples was measured with BCA protein assay kit (23225, Pierce Biotechnology, Rockford, IL, USA). Ca content of the cells was normalized to protein content and expressed as mg/mg protein. The observer who performed all the Ca measurements was blinded to the group assignment.

Chowdhury A., Balogh E., Ababneh H., Tóth A, & Jeney V. (2022). Activation of Nrf2/HO-1 Antioxidant Pathway by Heme Attenuates Calcification of Human Lens Epithelial Cells. Pharmaceuticals, 15(5), 493.

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Calcium Content Determination in Cells

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Cells grown in 96-well plates were washed twice with PBS and decalcified with 0.6 mol/L HCl for 30 minutes. Ca content of the supernatants was determined by the QuantiChrome Calcium Assay Kit (Gentaur, Kampenhout Belgium) as previously described [33] .

Balogh E., Tóth A., Tolnai E., Bodó T., Bányai E., Szabó D.J., Petrovski G, & Jeney V. (2016). Osteogenic differentiation of human lens epithelial cells might contribute to lens calcification. Biochimica et biophysica acta, 1862(9).

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