Cetylpyridinium chloride

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany, China, Switzerland, Japan, Austria

Cetylpyridinium chloride is a chemical compound that serves as a cationic surfactant. It is commonly used in various industrial and pharmaceutical applications.

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Lab products found in correlation

Alizarin red s, by Merck Group (77 mentions) Alizarin red, by Merck Group (40 mentions) Dexamethasone (dex), by Merck Group (31 mentions) Ascorbic acid, by Merck Group (22 mentions) Alizarin red solution, by Merck Group (21 mentions) β glycerophosphate, by Merck Group (21 mentions) Alizarin red s solution, by Merck Group (17 mentions) Paraformaldehyde (pfa), by Merck Group (15 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (13 mentions) Alizarin red s ar, by Merck Group (11 mentions) Ars solution, by Merck Group (9 mentions) β glycerol phosphate, by Merck Group (9 mentions) L ascorbic acid 2 phosphate, by Merck Group (8 mentions) Sodium phosphate, by Merck Group (6 mentions) Triton x 100, by Merck Group (6 mentions) Alizarin red staining, by Cyagen (6 mentions) L ascorbic acid, by Merck Group (6 mentions) Elx808, by Agilent Technologies (5 mentions) Microplate reader, by Thermo Fisher Scientific (5 mentions) Microplate reader, by Agilent Technologies (5 mentions)

Spelling variants of this product

Cetylpyridinium chloride (CPC) (50 citations) Cetylpyridinium chloride monohydrate (32 citations) Cetylpyridinium chloride solution (31 citations) Cetylpyridinium (7 citations) Cetylpyridinium chloride monohydrate solution (4 citations) Cetylpyridinium chloride (CPC) solution (3 citations) Cetylpyridinium chloride monohydrate (CPC, (2 citations) Cetylpyridinium chloride buffer (2 citations)

431 protocols using cetylpyridinium chloride

Roxadustat Enhances Osteogenic Differentiation

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Cells were cultured in osteogenic medium with or without roxadustat. ALP staining was performed at day 7 of roxadustat treatment. We fixed cells with 4% paraformaldehyde and stained them using an ALP staining kit (Beyotime, China) according to the manufacturer’s protocol. Cells were stained for 15 min at room temperature, and the staining was stopped by washing with distilled water three times. The integrated staining intensity was calculated with Image-Pro Plus software (IPP, Media Cybernetics, USA).
The matrix mineralization of cells was determined by Alizarin Red S staining at day 14 of roxadustat treatment. The cells were fixed with 4% paraformaldehyde and stained with 1% Alizarin Red S (Solarbio, China), pH 4.2, for 10 min at room temperature. Then, the cells were washed with distilled water three times to eliminate nonspecific staining. To quantify extracellular matrix mineralization, the bound stain was eluted with 10% (w/v) cetylpyridinium chloride (Sigma-Aldrich, St. Louis, MO, USA) in water at room temperature for 40 min followed by dilution with the same volume of 10% (w/v) cetylpyridinium chloride [18 (link)] and then quantified by measuring absorbance at 570 nm.

Li L., Li A., Zhu L., Gan L, & Zuo L. (2022). Roxadustat promotes osteoblast differentiation and prevents estrogen deficiency-induced bone loss by stabilizing HIF-1α and activating the Wnt/β-catenin signaling pathway. Journal of Orthopaedic Surgery and Research, 17, 286.

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Quantification of Mineralized Matrix

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After 21 days of co‐culture, the samples were rinsed three times with PBS, then fixed in 4% paraformaldehyde for 30 min, and finally the fixed samples were stained with ARS dye for 30 min. The stained samples were rinsed three times with deionized water to remove excess ARS dye. Images of the stained samples were obtained using fluorescence inverted microscopy. For further quantitative analysis of calcium deposition, the stained samples were reacted with 10% cetylpyridinium chloride (Sigma‐Aldrich, USA) for 1 h at room temperature, and the supernatant was taken in a 96‐well plate, and the absorbance at 562 nm was measured by UV–visible spectrophotometer (Perkin Elmer, USA).

Wang Y., Hu Y., Lan S., Chen Z., Zhang Y., Guo X., Cai L, & Li J. (2023). A Recombinant Parathyroid Hormone‐Related Peptide Locally Applied in Osteoporotic Bone Defect. Advanced Science, 10(22), 2300516.

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Osteogenic Differentiation of SHED Cells

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A SHED cell suspension (2.0×10⁴ cells/well) was seeded in culture dishes containing the different scaffolds and was cultured for 21 days in ODM (n = 3). The scaffolds were then washed with PBS (Sigma-Aldrich), fixed in 10% formaldehyde solution for 10 min and immersed in an aqueous solution with 1 vol% hydroxyl ammonium and 1 wt% Alizarin Red for 3 min. After several washes to remove excess stain, the scaffolds were dried, and the dye was desorbed from the scaffolds using 10% cetylpyridinium chloride (Sigma-Aldrich) for 1 h [33 (link)]. The absorbance of the Alizarin Red/calcium complex was measured at 570 nm for scaffolds in the presence or absence of cells. The initial mineral content of the scaffold composition was subtracted from the final mineral content.

Gonçalves F., de Moraes M.S., Ferreira L.B., Carreira A.C., Kossugue P.M., Boaro L.C., Bentini R., Garcia C.R., Sogayar M.C., Arana-Chavez V.E, & Catalani L.H. (2016). Combination of Bioactive Polymeric Membranes and Stem Cells for Periodontal Regeneration: In Vitro and In Vivo Analyses. PLoS ONE, 11(3), e0152412.

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Validating VC Cellular Model with Pi

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For the validation of the VC cellular model, primary RVSMCs were seeded in a 12-well plate. When cells reached about 80% confluence, 2 mM Pi was treated for 6 h, 3 days, and 6 days, respectively. For the confirmation of the effects of circSamd4a knockdown or its overexpression, A10 cells were seeded in a 24-well plate. After a day, cells were treated with either siRNA or overexpression vector. Then, 4 mM Pi was treated for an additional 3 days. For the ARS staining, cells were washed with PBS and fixed with 10% formalin at room temperature for 1 h. After removing 10% formalin, cells were washed three times with distilled water. When the samples were exposed to air and dried, 40 mM ARS solution (pH 4.2) (Sigma) was treated, and the samples were incubated overnight with rotation at room temperature. After removing the ARS solution, samples were washed with distilled water and PBS.
For ARS staining quantification, 300 μL 10% cetylpyridinium chloride (Sigma) was added to samples and incubated for 30 min. Then 200 μL dissolved solution was used to measure their absorbance at 562 nm by utilizing a microplate spectrophotometer (BioTek Instruments).

Ryu J., Kwon D.H., Choe N., Shin S., Jeong G., Lim Y.H., Kim J., Park W.J., Kook H, & Kim Y.K. (2019). Characterization of Circular RNAs in Vascular Smooth Muscle Cells with Vascular Calcification. Molecular Therapy. Nucleic Acids, 19, 31-41.

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Quantifying ECM Calcification Levels

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The extend of ECM calcification correlates with the grade of ECM maturation. The cells were fixed with 70% ethanol (Serva Electrophoresis, Heidelberg, Germany) in aqua dest. for 20 min at 6 °C after the CCM was discarded, and the cells were rinsed with DPBS beforehand. The fixation solution was removed, and the cells were once more washed with aqua dest. before 0.5% Alizarin Red S (Waldeck GmbH & Co. KG, Muenster, Germany) in aqua dest. was added for 10 min at room temperature. The supernatant of the staining solution was removed, and the cells underwent another washing procedure with aqua dest. to ensure only the bound stains remained. For quantification, the colored calcium was dissolved by adding 10% cetylpyridiniumchloride (Sigma-Aldrich) solution containing 10 mM sodiumdihydrogenphosphate (AppliChem, Darmstadt, Germany). Optical density was measured with a microplate reader (Autobio PHOmo) at 570 nm. For the quantitative determination of bound Alizarin Red S, a standard dilution series ranging from 90 µg down to 0 µg Alizarin Red S solution was plotted.

Rehder F., Arango-Ospina M., Decker S., Saur M., Kunisch E., Moghaddam A., Renkawitz T., Boccaccini A.R, & Westhauser F. (2024). The Addition of Zinc to the ICIE16-Bioactive Glass Composition Enhances Osteogenic Differentiation and Matrix Formation of Human Bone Marrow-Derived Mesenchymal Stromal Cells. Biomimetics, 9(1), 53.

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Quantification of Osteogenic Mineralization

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Following the 15-day culture period, the cells underwent PBS (phosphate-buffered saline) washing and fixation with 10% formaldehyde at room temperature for 15 min. Subsequently, the cells were rinsed thrice with ddH₂O and stained with a 40 mM alizarin red solution (Sigma-Aldrich, Merck Group) at room temperature for 30 min. After staining, the cells were washed four times with ddH₂O. For quantification, the stained mineralized nodules were subjected to a 30 min incubation with 10% cetylpyridinium chloride (Sigma-Aldrich). The resulting solutions were collected, and absorbance at 562 nm was measured using a Tecan Infinite M200 microplate reader. Sample quantification was performed based on the alizarin red standard curve. The inclusion of alizarin red staining, a widely accepted method for assessing calcium deposition, complemented the quantitative measures by providing a qualitative and visual evaluation of mineralization [68 (link)].

Harahap I.A., Olejnik A., Kowalska K, & Suliburska J. (2024). Effects of Daidzein, Tempeh, and a Probiotic Digested in an Artificial Gastrointestinal Tract on Calcium Deposition in Human Osteoblast-like Saos-2 Cells. International Journal of Molecular Sciences, 25(2), 1008.

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Optimal hADSC-Exosomes for Osteogenesis

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To determine the optimal time-span of extracted hADSC-Exos during hADSCs osteogenic differentiation, the hADSCs of passage 4 were seeded into the 24-well plates, and was cultured in PM. After getting 70–80% confluence, the culture medium was replaced by PM containing exosomes (Exo^0d, Exo^1−14d and Exo^15−28d) from different time-span of hADSCs osteogenesis induction. hADSCs cultured in exosomes-free PM and OM were set as negative and positive controls, respectively.
The mineralized deposits were evaluated by ARS after incubation for 21 days. After fixation by 4% paraformaldehyde for 30 min, the cells were washed three times with PBS before staining with Alizarin red S for 5 min, and then the samples were washed again with PBS. For ARS quantitation the mineralized deposits were dissolved in 10% cetylpyridinium chloride (Sigma, USA) in dark for 30 min at room temperature. The solution was transferred to a 96-well plate with 100 μL/well. The optical density (OD) values was measured at 562 nm using a microplate reader (BioTek EL808, USA).

Zhu M., Liu Y., Qin H., Tong S., Sun Q., Wang T., Zhang H., Cui M, & Guo S. (2020). Osteogenically-induced exosomes stimulate osteogenesis of human adipose-derived stem cells. Cell and Tissue Banking, 22(1), 77-91.

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Resveratrol-Induced Osteogenic Differentiation

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BMSCs were seeded into 24-well plates and cultured with osteogenic differentiation medium (Cyagen Biosciences, USA) after resveratrol treatment. After 3 days or 7 days cell culture, cells were fixed with 4% formaldehyde solution (Boster Biological Technology co. ltd, China) for 30 min and washed twice with PBS. Then cells were stained with alizarin red dye (Cyagen Biosciences, USA) for 5 min, washed twice with PBS, and imaged in the light field to observe the calcium nodules. To quantitatively analyze the calcification, 2 mL cetylpyridinium chloride (Sigma-Aldrich, USA) was added into each well to dissolve the calcium nodules. The absorbance at 562 nm was measured.

Zhou T., Yan Y., Zhao C., Xu Y., Wang Q, & Xu N. (2019). Resveratrol improves osteogenic differentiation of senescent bone mesenchymal stem cells through inhibiting endogenous reactive oxygen species production via AMPK activation. Redox Report : Communications in Free Radical Research, 24(1), 62-69.

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Quantifying Mineralization of mMSCs

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The mineralization of mMSCs, which were differentiation-induced for 21 days, was determined by ARS staining. The cells were rinsed twice using PBS, fixed in 4% paraformaldehyde at 4 °C for 30 min, and washed using deionized water. Cell staining was performed using 40 mM ARS solution (pH 4.2; Sigma) for 30 min at room temperature. Subsequently, the cells were rinsed three times with double-distilled water and washed using PBS for 15 min to reduce nonspecific ARS staining. The stained cells were observed under a microscope. To quantify the mineralization, the stained layers were solubilized by the addition of 10% cetylpyridinium chloride (pH 7.0; Sigma) as previously described11 (link). Dye absorbance was determined at 570 nm. Six wells were analyzed per experiment and the experiments performed in triplicate.

Wang J., Tao Y., Ping Z., Zhang W., Hu X., Wang Y., Wang L., Shi J., Wu X., Yang H., Xu Y, & Geng D. (2016). Icariin attenuates titanium-particle inhibition of bone formation by activating the Wnt/β-catenin signaling pathway in vivo and in vitro. Scientific Reports, 6, 23827.

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Osteogenic Differentiation Quantification

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Cells were seeded in a 12-well plate at a concentration of 5 × 10³ cells /well and cultured with α-MEM supplemented with 10% FBS until 80% confluency. Cells in differentiation group were incubated with osteogenic induction medium consisted of α-MEM supplemented with 10% FBS, 0.1 µM dexamethasone, 10 mM β-glycerophosphate (Sinopharm Chemical Reagent Ltd., Shanghai, China) and 200 µM ascorbic acid (Sigma, St. Louis, MO, USA). Cells in control group were cultured with α-MEM supplemented with 10% FBS. Medium was changed every three days. After incubation for three weeks, cells were stained by Alizarin red (Chroma-Schmidt GmbH, Köngen, Germany) and observed under an inverted microscope. Then osteogenic differentiation was quantified by an enzyme immunoassay analyzer at 570 nm after the solubilization with 10% cetylpyridinium chloride (Sigma, St. Louis, MO, USA) for 10 min at room temperature.

Zhang F., Ren H., Shao X., Zhuang C., Chen Y, & Qi N. (2017). Preservation media, durations and cell concentrations of short-term storage affect key features of human adipose-derived mesenchymal stem cells for therapeutic application. PeerJ, 5, e3301.

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