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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
成骨细胞染色试剂盒(茜素红S)(Osteoblast Staining Kit (Alizarin Red S)),也称骨细胞矿化结节染色试剂盒(茜素红S法)和 成软骨鉴定试剂盒(茜素红),是一种基于茜素红S螯合钙离子(Ca²⁺)生成橙红色复合物,以检测成骨细胞矿化结节的染色试剂盒。
人体和动物的骨组织在生命过程中不断地进行着重建,骨重建过程包括骨的分解吸收与新骨的形成。破骨细胞(osteoclasts)负责骨分解与吸收,而成骨细胞(osteoblasts,OB)是骨形成的主要功能细胞,负责新骨形成。在成骨分化过程中,其基本的生物学特征是骨基质合成、分泌、矿化及成熟。成骨细胞首先合成I型胶原(collagen-I,COL-I)、骨钙素(osteocalcin,OC)、骨桥蛋白(osteopontin,OPN)等胞外基质,并通过基质小泡释放钙离子和碱性磷酸酶(alkalinephosphatase,ALP)等酶类物质,钙离子在碱性磷酸酶作用下沉淀在胶原纤维丝上,完成基质矿化过程,最终形成骨组织。I型胶原从增殖期开始表达,基质合成期达巅峰;碱性磷酸酶是成骨细胞分化的早期标志物,骨基质合成期开始出现,矿化期达巅峰;骨钙素是成骨细胞分化成熟的标志,一般矿化早期开始表达,矿化结节成熟后达巅峰。矿化结节是成骨细胞分化成熟的标志,同时也是成骨细胞行使成骨功能的主要形态学特征,观察成骨细胞的矿化结节是研究成骨细胞分化的常用技术方法之一。
茜素红S染色是一种常用的染色方法,广泛用于成骨细胞分化、骨细胞或组织病理生理的研究。茜素红S(AlizarinRedS,简称ARS),别名茜素磺酸钠、茜素胭脂红,CAS号为130-22-3,分子式为C₁₄H₇NaO₇S,分子量为342.25。茜素红S是一种蒽醌类衍生物,能和多种金属离子螯合生成稳定的橙红色或深红色的复合物,是一种良好的金属指示剂,也可作为吸光度法测定金属离子的显色剂,还可作为酸碱指示剂,或用于极谱法测定金属离子。在组织学和组织病理学中,由于茜素红S可与碳酸钙或磷酸钙中的钙盐螯合形成橙红色复合物,而常用于染色或定位组织中的钙质沉积。
| 项目号 | Components | 50T | Storage |
| O1506007A | 固定液 | 50 mL | 2-8℃ |
| O1506007B | 茜素红S染色液 | 50 mL | 2-8℃. Store in the dark |
使用说明:
1.样品处理。
a.细胞诱导分化之后,去除培养液,用PBS洗涤1次。
b.固定液固定20分钟。
c.PBS洗涤3次。
2.茜素红S染色液染色。
a.加入适量茜素红S染色液,均匀覆盖细胞即可,室温染色30分钟。注:请根据实际染色效果调整染色时间。
b.蒸馏水充分洗涤,随后即可在显微镜下观察和拍照。
注意事项:
茜素红S染色液的染色的最佳时间与样品钙离子的含量有关,一般情况下染色时间为30分钟,可根据实际染色效果酌情增加或减少染色时间。
Osteogenesis Assay Kit (Alizarin Red S), Also known as the Alizarin Red S Staining Kit for Osteogenesis and the Osteogenesis Assay Kit, this kit is a staining tool designed to detect mineralized nodules of osteoblasts. Its working principle is based on the ability of Alizarin Red S to chelate calcium ions (Ca²⁺), forming an orange-red complex.
In the life process of humans and animals, bone tissue undergoes continuous remodeling. The bone remodeling process includes the resorption (breakdown) of old bone and the formation of new bone. Osteoclasts are responsible for bone resorption, while osteoblasts (OBs) are the primary functional cells for bone formation, tasked with generating new bone. During osteogenic differentiation, the basic biological characteristics of osteoblasts include the synthesis, secretion, mineralization, and maturation of the bone matrix. First, osteoblasts synthesize extracellular matrix components such as type I collagen (collagen-I, COL-I), osteocalcin (OC), and osteopontin (OPN). They then release calcium ions and enzymes like alkaline phosphatase (ALP) through matrix vesicles. Under the action of alkaline phosphatase, calcium ions precipitate onto collagen fibrils, completing the matrix mineralization process and eventually forming bone tissue. Type I collagen begins to be expressed during the proliferation phase and reaches its peak in the matrix synthesis phase. Alkaline phosphatase is an early marker of osteoblast differentiation; it appears in the matrix synthesis phase and peaks in the mineralization phase. Osteocalcin is a marker of osteoblast maturation, typically starting to be expressed in the early mineralization stage and peaking after the mineralized nodules mature. Mineralized nodules are not only a marker of osteoblast maturation but also the main morphological feature of osteoblasts exerting their bone-forming function. Observing the mineralized nodules of osteoblasts is one of the common technical methods for studying osteoblast differentiation.
Alizarin Red S staining is a widely used method in research on osteoblast differentiation and the pathophysiology of osteocytes or bone tissue. Alizarin Red S (abbreviated as ARS), also known as sodium alizarinsulfonate or alizarin carmine, has a CAS number of 130-22-3, a molecular formula of C₁₄H₇NaO₇S, and a molecular weight of 342.25. As an anthraquinone derivative, Alizarin Red S can chelate with various metal ions to form stable orange-red or crimson complexes. It serves multiple purposes: it is an excellent metal indicator, a chromogenic agent for the spectrophotometric determination of metal ions, an acid-base indicator, and a reagent for the polarographic determination of metal ions. In histology and histopathology, Alizarin Red S is commonly used to stain or localize calcium deposits in tissues, as it can chelate with calcium salts in calcium carbonate or calcium phosphate to form an orange-red complex.
| O1506007 | Components | 50T | Storage |
| O1506007A | Fixative | 50 mL | 2-8℃ |
| O1506007B | Alizarin Red S Staining Solution | 50 mL | 2-8℃. Store in the dark |
Instructions for Use:
1. Sample Preparation
a. After inducing cell differentiation, remove the culture medium and wash the cells once with PBS.
b. Fix the cells with fixative for 20 minutes.
c. Wash the cells three times with PBS.
2. Staining with Alizarin Red S Staining Solution
a. Add an appropriate amount of Alizarin Red S Staining Solution—just enough to cover the cells evenly—and stain at room temperature for 30 minutes.
Note: Adjust the staining time according to the actual staining effect.
b. Wash thoroughly with distilled water, then observe and take photos under a microscope.
Precautions:
The optimal staining time for Alizarin Red S Staining Solution is related to the calcium ion content in the sample. Under normal circumstances, the staining time is 30 minutes, but it can be increased or decreased as appropriate based on the actual staining effect.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | O1506007 |
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