计算溶液所需的质量、体积或浓度。
BioReagent,生物染色剂,用于显微镜 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存,避光,室温,-20°C储存。低温运输 。请查阅批次 COA 获取详细规格。
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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。
阿利新蓝又称爱先蓝或阿尔辛蓝等,是一种大分子共轭染料,类铜钛花青染料,最初用于纺织纤维染色,这种阳离子染料与酸性基团结合,也即阿利新蓝与组织内含有的阴离子基团如羧基和硫酸根形成不溶性复合物。组织学染色时推荐使用阿利新蓝8GX和8GS,阿利新蓝使碳水化合物着色的确切机制不明,普遍认为是阳离子的异硫脲基通过静电与组织内的多聚阴离子相连,不能沉淀糖原。硫酸软骨素、硫酸皮肤素、硫酸乙酰肝素和透明质酸的硫酸根或羧基在pH值为2.5时电离,因此带有一个负电荷,这导致结缔组织和软骨中的蛋白多糖/透明质酸以及上皮酸性黏蛋白(如大肠的唾液黏蛋白和硫黏蛋白)与阿利新蓝在pH值为2.5时发生反应,中性黏蛋白(如胃黏膜和Brunner腺体部位的中性黏蛋白)不能与阿利新蓝反应。
透明质酸染色液采用透明质酸酶裂解并联合阿利新蓝染色,其原理在于用透明质酸酶可以裂解透明质酸和硫酸软骨素中的糖苷键,在阿利新蓝染色前,用透明质酸酶做标本预处理。与未处理的切片相比,若处理过的切片不着色,则表明组织中存在透明质酸或硫酸软骨素;若预处理没有产生影响,则说明组织中不含透明质酸。虽然缺乏特异性,但经透明质酸酶处理后仍呈阴性可排除透明质酸存在的可能性。该试剂仅用于科研领域,不适用于临床诊断或其他用途。
产品组分及储存条件:
| H1518548 | Component | 2×50mL | Storage |
| H1518548A | 透明质酸酶溶液 | 25mL | -20℃. Store in the dark. |
| H1518548B | 磷酸盐缓冲液 | 25mL | RT |
| H1518548C | 阿尔辛蓝染色液 | 50mL | 2-8℃. Store in the dark. |
自备材料:
去离子水、系列乙醇/二甲苯或环保浸蜡脱蜡透明液、中性树胶
操作步骤(仅供参考):
1、取2张阳性对照片和2张实验片,二甲苯或脱蜡透明液脱蜡,通过梯度乙醇后,再入去离子水水化。
2、取1张阳性对照片和1张实验片用透明质酸酶溶液处理,37°C孵育3h;取另外的1张阳性对照片和1张实验片用磷酸盐缓冲液处理,37°C孵育3h。
3、流水冲洗5min。
4、入阿尔辛蓝染色液染色30min,流水冲洗5min。
5、梯度乙醇脱水,二甲苯或脱蜡透明液透明,中性树胶封片。
染色结果:
透明质酸和(或)硫酸软骨素(未经酶处理):蓝色
透明质酸和(或)硫酸软骨素(经酶处理):不着色
注意事项:
1、需使用阳性对照片以检测酶的活性。
2、为了您的安全和健康,请穿实验服并戴一次性手套操作。
Alcian Blue, also known as Alcian Blue 8GX, is a macromolecular conjugated dye belonging to copper phthalocyanine dyes. It was originally used for dyeing textile fibers. This cationic dye binds to acidic groups; that is, Alcian Blue forms insoluble complexes with anionic groups in tissues, such as carboxyl and sulfate groups. Alcian Blue 8GX and 8GS are recommended for histological staining. The exact mechanism by which Alcian Blue stains carbohydrates is unclear. It is generally believed that the cationic isothiouronium groups bind to polyanions in tissues electrostatically, and it does not precipitate glycogen. Sulfate or carboxyl groups of chondroitin sulfate, dermatan sulfate, heparan sulfate, and hyaluronic acid ionize at pH 2.5, thus carrying a negative charge. This causes proteoglycans/hyaluronic acid in connective tissue and cartilage, as well as epithelial acid mucins (e.g., sialomucins and sulfomucins of the large intestine), to react with Alcian Blue at pH 2.5. Neutral mucins (e.g., those in the gastric mucosa and Brunner’s glands) do not react with Alcian Blue.
Hyaluronic Acid Staining Solution uses hyaluronidase digestion combined with Alcian Blue staining. The principle is that hyaluronidase cleaves the glycosidic bonds in hyaluronic acid and chondroitin sulfate. Specimens are pretreated with hyaluronidase before Alcian Blue staining. Compared with untreated sections, if the treated sections show no staining, hyaluronic acid or chondroitin sulfate is present in the tissue. If the pretreatment has no effect, the tissue does not contain hyaluronic acid. Although lacking absolute specificity, a negative result after hyaluronidase treatment can rule out the presence of hyaluronic acid. This reagent is for research use only. Not for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
| H1518548 | Component | 2×50mL | Storage |
| H1518548A | HAase Solution | 25mL | -20℃. Store in the dark. |
| H1518548B | PB Buffer | 25mL | RT |
| H1518548C | Alcian Blue Stain | 50mL | 2-8℃. Store in the dark. |
Materials Required:
Deionized water, graded ethanol / xylene or eco‑friendly dewaxing and clearing agent, neutral balsam
Protocol (For Reference Only):
1. Take 2 positive control slides and 2 test slides. Deparaffinize with xylene or dewaxing and clearing agent, rehydrate through graded ethanol, then place in deionized water.
2. Incubate 1 positive control slide and 1 test slide with HAase Solution at 37 °C for 3 h. Incubate the other 1 positive control slide and 1 test slide with PB Buffer at 37 °C for 3 h.
3. Rinse in running water for 5 min.
4. Stain in Alcian Blue Stain for 30 min, then rinse in running water for 5 min.
5. Dehydrate in graded ethanol, clear in xylene or dewaxing and clearing agent, and mount with neutral balsam.
Staining Results:
Hyaluronic acid and/or chondroitin sulfate (without enzyme treatment): Blue
Hyaluronic acid and/or chondroitin sulfate (with enzyme treatment): Negative / No staining
Precautions:
1. Positive control slides must be used to verify enzyme activity.
2. For your safety and health, wear a lab coat and disposable gloves during operation.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | H1518548 |
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