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Useful for staining calcified nodule

Calcified Nodule Staining Kit

Background

Alkaline phosphatase, osteopontin, and osteocalcin are expressed during the differentiation of bone marrow cells into mature osteoblasts and osteoblasts form calcium tubercle. Alizarin Red S, an anthraquinone derivative, has been used to identify Calcified nodule, because Calcium forms an Alizarin Red S-calcium complex, and the products turn (became) red. Calcified Nodule Staining kit (PMC-AK21-COS) contains Substrate-containing Buffer composed mainly of Alizarin Red S and Chromogenic Substrate, and provides a convenient system for staining calcified nodule with ease

Kit component

Component Quantity Storage
Substrate-containing Buffer (×100) 60 mL 4°C
Chromogenic Substrate 10 vials 4°C

Protocol (24well-plate format)

1. Remove culture medium. Wash each well with 1 mL of PBS. (× 3 times)
2. Remove PBS, and fix cells with 500 uL of methanol per well at room temperature for 30 minutes.
  * Methanol should be cooled at -20 °C before use.
3. Remove methanol. Wash each well with 1 mL of distilled water. (× 3 times)
4. Add one vial of Chromogenic Substrate to 10 mL of Substrate-containing Buffer and mix, and then dispense 400 μL of the working solution to each well.
  * This working solution should not be stored. Ready to use.
5. Incubate the plate at room temperature for 5 minutes
6. Wash each well with 1 mL of Substrate-containing Buffer until washing buffer become transparent.

Application example

It's applicable samples regardless of animal species.

Cells detected by Calcified Nodule Staining kit

A: 3T3-E1 cells are cultured using Osteogenesis Culture for Osteogenesis Culture kit (Mouse) (Code No. PMC- OGCMO-COS) (35 mm dish)
B: Cells in Osteogenesis Culture Kit (Mouse) (PMC-OGC11-COS) (24well plate) 

Product List

Product Name Cat# Quantity Price

Calcified nodule Staining kit

PMC-AK21-COS 1SET

¥ 47,000
$ 627
€ 470

Citations
  • • “L.T. Bang, et al. Effects of silicate and carbonate substitution on the properties of hydroxyapatite prepared by aqueous co-precipitation method. Materials and Design 87 (2015) 788–796 Article: S0264127515303191
  • • “R. Toita, et al. Modulation of the osteoconductive property and immune response of poly(ether ether ketone) by modification with calcium ions. Journal of Materials Chemistry B (2015) 3, 8738-8746 DOI: 10.1039/C5TB01679G
  • • “Maruyama K, et al. TRAF family member-associated NF-κB activator (TANK) is a negative regulator of osteoclastogenesis and bone formation. J Biol Chem. 2012 Aug 17;287(34):29114-24 PMID: 22773835
  • • “Mayu K, et al. Cell Proliferation Ability of Mouse Fibroblast-Like Cells and Osteoblast-Like Cells on a Ti-6Al-4V Alloy Film Produced by Selective Laser Melting. Materials Sciences and Applications, 2014, 5, 475-483 Article ID: 46462
  • • “Nakamura K, et al. ROCK inhibitor Y-27632 maintains the proliferation of confluent human mesenchymal stem cells. J Biol Chem. J Periodontal Res. 2014 Jun;49(3):363-70 PMID: 23834550
  • • “Taniguchi Y, et al. Accelerated Osteogenic Differentiation and Bone Formation on Zirconia with Surface Grooves Created with Fiber Laser Irradiation. J Biol Chem. J Periodontal Res. Clin Implant Dent Relat Res. 2015 Jul 14 PMID: 26179832

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.

Cell Assay Related Research Kit