BMSCs in different groups were cultured in six-well plates containing osteogenic differentiation medium (Cyagen, Suzhou, China). Media were changed every 3 days for 3 weeks. After rinsing, cells were fixed with 4% paraformaldehyde. Mineralized nodules were observed by staining with a 1% alizarin red solution (Cyagen, Suzhou, China) following manufacturer instructions [12 (link)]. ALP staining was used to investigate the osteogenesis of BMSCs after induction [18 (link)]. Staining images were recorded under an inverted microscope, and ImageJ was used to analyze the results.
Alizarin red solution
Manufactured by Cyagen
Sourced in China, United States
Alizarin red solution is a laboratory reagent used for the staining and visualization of calcium deposits. It is a red dye that binds to calcium ions, producing a characteristic red color. The solution is commonly used in histological and cell culture applications to identify the presence of mineralized structures.
Automatically generated - may contain errors
Lab products found in correlation
Osteogenic differentiation medium, by Cyagen (4 mentions)
Cetylpyridinium chloride, by Merck Group (4 mentions)
Dexamethasone (dex), by Cyagen (3 mentions)
Dexamethasone (dex), by Merck Group (3 mentions)
6 well plate, by Thermo Fisher Scientific (3 mentions)
Cetylpyridinum chloride, by Merck Group (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Alizarin red, by Cyagen (1 mentions)
Accutase, by Thermo Fisher Scientific (1 mentions)
Osteogenic differentiation medium, by Merck Group (1 mentions)
Cetylpyridine, by Merck Group (1 mentions)
Alp detection kit, by Beyotime (1 mentions)
Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Bcip nbt alkaline phosphatase color reagent kit, by Beyotime (1 mentions)
Bca protein assay kit, by Nanjing Jiancheng (1 mentions)
Oil red o solution, by Solarbio (1 mentions)
N2 dmi8, by Leica (1 mentions)
β glycerol phosphate, by Cyagen (1 mentions)
Ascorbic acid, by Cyagen (1 mentions)
27 protocols using alizarin red solution
1
Osteogenic Differentiation of BMSCs
2
Osteogenic Differentiation of HPDLSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HPDLSCs (2 × 105 cells/well) were cultured in 6-well plates containing osteogenic differentiation medium (Cyagen, Suzhou, China). The medium was refreshed every 3 days until the 21st day. Cells were fixed with 4% paraformaldehyde. Mineralized nodules were observed by staining with the 1% Alizarin red solution (Cyagen, Suzhou, China). The activity of alkaline phosphatase (ALP) was analyzed using an Alkaline Phosphatase Staining Kit (MKbio, Shanghai, China). Staining images were acquired using an inverted microscope, and ImageJ was used to analyze the images.
3
Osteogenic Differentiation of BMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMSCs in different groups were cultured in six-well plates containing osteogenic differentiation medium (Cyagen, Suzhou, China). Media were changed every 3 days for 3 weeks. After rinsing, cells were xed with 4% paraformaldehyde. Mineralized nodules were observed by staining with a 1% alizarin red solution (Cyagen, Suzhou, China) following manufacturer instructions [12] . ALP staining was used to investigate the osteogenesis of BMSCs after induction [18] . Staining images were recorded under an inverted microscope, and ImageJ was used to analyze the results.
4
Alizarin Red-based ECM Mineralization Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ECM mineralization of C3H10T1/2 was analyzed with Alizarin Red staining. After culturing in the extracts for 7 and 14 days, the cells were fixed with 75% ethanol for 1 h and subsequently stained with 2% Alizarin Red solution (Cyagen, China). Images were taken by an optical microscope (Olympus, Germany). The bonded dyes were dissolved in 10 mM sodium phosphate aqueous solution containing cetylpyridinum chloride (Sigma-Aldrich, USA) and the absorbance was measured at 540 nm with a microplate reader (Bio-Tek, USA).
5
Osteogenic Induction and Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow mesenchymal stem cells that underwent osteogenic induction were fixed with 4% paraformaldehyde for 10 minutes. The ALP staining was conducted with the BCIP/NBT regent kit (Beyotime), and Alizarin Red staining, cells were stained with Alizarin Red solution (Cyagen Biosciences). All procedures were carried according to the manufacture's protocols.
6
Osteoblast Differentiation from H9-NCCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For MSCs differentiation, H9-NCCs were treated with Accutase (Thermal Fisher, Carlsbad, CA) and then plated on 0.1% gelatin-coated plate with MSC differentiation medium as described 42 . MSCs were subsequently induced into osteoblast by culturing in osteogenic differentiation medium (Cyagen, Santa Clara, CA) and stained by Alizarin Red solution (Cyagen) after 2 weeks of culture.
7
Osteogenic Differentiation of rBMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The osteogenic differentiation of rBMSCs was evaluated using alkaline phosphatase (ALP) and Alizarin Red (AR) staining. The rBMSCs were seeded into 48-well plates at a density of 1 × 104 cells/well. After the cells reached about 80% confluence, the medium was replaced with osteogenic differentiation medium (Sigma-Aldrich, USA) containing different scaffold extracts and supplemented with 10 mM404 β-glycerophosphate, 50 mM ascorbic acid, and 100 nM dexamethasone. As mentioned earlier[18 (link)], ALP staining was performed on the day 7, and ALP activity was determined using an ALP detection kit (Beyotime Biotechnology, China) according to the manufacturer’s instructions. To evaluate the formation of calcium nodules, rBMSCs were stained with Alizarin Red solution (Cyagen, USA) 21 days after osteogenic induction. After photography and recording, the mineralized nodules were dissolved in 10% cetylpyridine (Sigma-Aldrich, USA), and the absorbance at 562 nm was measured for semi-quantitative analysis.
8
Alizarin Red Staining of Mineralized Matrix
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The formation of mineralized matrix nodules was determined using ARS on the 21st day after the hBMSCs were seeded in the 24-well plate with osteogenic induction medium. The cells were washed with PBS for three times, fixed with 4% paraformaldehyde, and then stained with Alizarin red solution (Cyagen Biosciences, Santa Clara, CA, USA) for 15 minutes, according to manufacturer’s instructions.
9
Polylactic Acid-based Biomaterial Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polylactic acid (PLLA, Mw = 200000, PDI = 1.5, CAS: 33135-50-1) was provided by Jinan Daigang Biomaterials Co., Ltd. Cholesteryl oleyl carbonate (COC, CAS: 17110-51-9) was purchased from Tokyo Chemical Industry. Cholesteryl pelargonat (CP, CAS: 1182-66-7) was purchased from Sigma-Aldrich. Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and penicillin–streptomycin solution were purchased from Gibco BRL. The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies Inc. The AO-EB Double Staining Kit was purchased from Beijing Solarbio Science and Technology Co., Ltd. Rhodamine phalloidin and 4′, 6-dimidyl-2-phenylindole (DAP, CAS:28718-90-3) were purchased from Molecular Probes, United States. Osteoblastic induction medium and Alizarin Red solution were purchased from Cyagen Biosciences Inc. Fluorescein isothiocyanate conjugated anti-mouse CD206 (FITC-CD206) and Allophycocyanin conjugated anti-mouse CD80 (APC-CD80) were purchased from Dakewe Biotech Co., Ltd. The BCIP/NBT alkaline phosphatase color reagent kit was purchased from Beyotime Biotechnology Co., Ltd. ALP and the BCA protein assay kit was purchased from Nanjing Jiancheng Bioengineering Institute. All cells were obtained from the American Type Culture Collection (ATCC). All other reagents were purchased from Guangzhou Reagent Co., Ltd., analytical grade.
10
Multilineage Differentiation of Bone Marrow Stromal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In osteogenic differentiation, BMSCs were seeded in 48-well plates as 2 × 104/well, and the medium was replaced with osteogenic induction medium (Minimum Essential Medium α +10% fetal bovine serum (FBS) + 0.1% dual antibiotics +0.1 μmol/L dexamethasone +50 μmol/L ascorbic acid +10 mmol/L β-glycerol phosphate) (Cyagen, China) at 48h. After culturing for 14 days, BMSCs were stained with alizarin red solution (Cyagen, China) and observed (Leica, N2-DMi8, Germany). In adipogenic differentiation, BMSCs were cultured with adipogenic differentiation induction medium (10% FBS +1% glutamine +0.2% insulin +0.1% IBMX +0.1% rosiglitazone +2.5% dexamethasone) (Cyagen, China). Oil red O solution (Solarbio, China) was added to stain the lipid droplet at 21 days. As for chondroblast differentiation, BMSCs were cultured with chondrogenic induction medium (DMEM medium + 0.1 μmol/L dexamethasone +1.25 mg/mL bovine serum albumin +1 mmol/L sodium pyruvate + ITS +10 ng/mL TGF-β+37.5 μg/mL vitamin C+ 1 ng/mL β-FGF) (Cyagen, China) for 14 days, and the cartilage balls were sliced and stained with cartilage staining solution (Cyagen, China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!