Alcian Blue 8GX
(Synonyms: 阿尔新蓝8GX) 目录号 : GC67217
Alcian Blue 8GX 是一种阳离子铜酞菁染料，众所周知可与糖蛋白和葡糖胺聚糖结合。
Cas No.:33864-99-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
本方案仅提供一个指导，请根据您的具体需要进行修改。
1、 制备1 -10mg/mL的Alcian Blue 8GX染色液：向ddH2O中加入0.6mL冰醋酸，使最终体积为20 mL。然后加入20-200 mg固体Alcian Blue 8GX，充分溶解后使用0.22μm滤膜过滤除菌。
注意：配置好的染液4℃避光保存，建议在一个月内用完。
2、细胞悬浮染色(以6 孔板为例)
（1）悬浮细胞经1000g离心3-5min。弃去上清液，使用PBS清洗两次，每次5分钟。
（2）贴壁细胞使用PBS清洗两次，加入胰酶消化细胞，消化完成后经1000g离心3-5min。
（3）加入1mL染料工作液重悬细胞，室温避光孵育30-60min分钟，不同细胞最佳培养时间不同。
（4）孵育结束后，经1000g离心5分钟，去除上清液，加入PBS清洗2-3次，每次5分钟。
（5）使用无血清细胞培养基或PBS重悬细胞，通过显微镜进行观察。
3、细胞贴壁染色
（1）在无菌盖玻片上培养贴壁细胞。
（2）从培养基中移走盖玻片，吸出过量的培养基，将盖玻片放在潮湿的环境中。
（3）从盖玻片的一角加入100μL染料工作液，轻轻晃动使染料均匀覆盖所有细胞，室温避光孵育30-60min分钟。
（4）吸弃染料工作液，使用培养液清洗盖玻片2~3次，每次5分钟。
4、显微镜检测：使用光学显微镜对染色细胞进行观察。
注意事项：为了您的安全和健康，请穿实验服并戴一次性手套操作。
References:
[1]. Hiroki Ueharu,et. Isolation and Culture of Cranial Neural Crest Cells from the First Branchial Arch of Mice. 2022 Apr 5;12(7):e4371. doi: 10.21769/BioProtoc.4371.
[2]. J Tas. The Alcian blue and combined Alcian blue--Safranin O staining of glycosaminoglycans studied in a model system and in mast cells. 1977 Mar;9(2):205-30. doi: 10.1007/BF01003632.
Alcian Blue 8GX is a cationic copper phthalocyanine dye well-known to bind to glycoproteins and to glucosaminoglycans[1].
[1]. Candiano G, et al. Widening and diversifying the proteome capture by combinatorial peptide ligand libraries via Alcian Blue dye binding. Anal Chem. 2015;87(9):4814-4820.
| Cas No. | 33864-99-2 | SDF | Download SDF |
| 别名 | 阿尔新蓝8GX | ||
| 分子式 | C56H68Cl4CuN16S4 | 分子量 | 1298.86 |
| 溶解度 | 储存条件 | Store at -20°C | |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 0.7699 mL | 3.8495 mL | 7.6991 mL |
| 5 mM | 0.154 mL | 0.7699 mL | 1.5398 mL |
| 10 mM | 0.077 mL | 0.385 mL | 0.7699 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Alcian blue pyridine variant--a superior alternative to Alcian Blue 8GX: staining performance and stability
Biotech Histochem 2000 May;75(3):147-50.PMID:10950177DOI:10.3109/10520290009066493.
We compared the staining performance, dye content, solubility, and visual absorption maximum of two batches of alcian blue pyridine variant and of five batches of Alcian Blue 8GX (C.I. 74240). Whenever possible, we also compared results to those obtained with the same dye batches produced at an earlier date to provide information concerning dye stability. Both alcian blue pyridine variant batches were of high dye content, stable, of satisfactory solubility, and performed well in both the routine Mowry mucin stain and in the critical electrolyte concentration (CEC) stain. Of the five Alcian Blue 8GX samples, some were also of appropriate dye content, were sufficiently stable, and gave good staining in the two procedures. Two batches, however, were unstable, and three batches were unsatisfactory in staining performance and solubility in the CEC stain. Consequently alcian blue pyridine variant is a superior substitute for Alcian Blue 8GX.
An alternative Alcian Blue dye variant for the evaluation of fetal cartilage
Birth Defects Res B Dev Reprod Toxicol 2007 Jun;80(3):171-6.PMID:17410541DOI:10.1002/bdrb.20109.
Background: The most comprehensive evaluation of vertebrate skeletal development involves the use of Alizarin Red S dye to stain ossified bone and various other dyes to stain cartilage. The dye used most widely to stain fetal cartilage in rodents and rabbits is Alcian Blue 8GX. However, the global supply of this specific dye has been exhausted. Several forms of the dye marketed as Alcian Blue 8GX are now available, although they are not synthesized via the original 8GX manufacturing process. Methods: One new Alcian Blue 8GX form and two Alcian Blue dye variants were evaluated in rats and rabbits using standard staining procedures. The staining quality of these dyes were evaluated relative to the original form of Alcian Blue 8GX based on cartilage uptake of the dye, clarity of the cartilaginous components, staining intensity of the dye, and overall readability of the specimens under stereomicroscopic evaluation. Results: Staining with the newer form of Alcian Blue 8GX resulted in poor staining quality. The Alcian Blue-Pyridine variant performed well, although staining intensity was less than optimal. The Alcian Blue-Tetrakis variant provided staining characteristics that were most similar to the original form of Alcian Blue 8GX. Conclusions: Alcian Blue-Tetrakis was markedly better in its ability to stain fetal cartilage than the newer form of Alcian Blue 8GX.
The quantitative determination of glycosaminoglycans in urine with Alcian Blue 8GX
Biochem J 1973 Feb;131(2):351-7.PMID:4269150DOI:10.1042/bj1310351.
1. The effect of MgCl(2) concentration on the interaction of Alcian Blue 8GX and glycosaminoglycans in the urine of patients with mucopolysaccharidosis was studied by using a new quantitative micro method for the measurement of Alcian Blue-glycosaminoglycan complexes. This provided a means of measuring the critical electrolyte concentrations of urinary glycosaminoglycans. 2. Theoretical considerations based on the preceding paper (Whiteman, 1973) and experimental evidence provided here show that Alcian Blue 8GX may be used for the direct quantitative determination of total urinary glycosaminoglycans. The method is simple, requires sample volumes of 50mul or less, and gives results comparable with those obtained by other more complicated methods.
Staining of interphase nuclei and mitotic figures in cultured cells with Alcian Blue 8GX
In Vitro 1982 May;18(5):456-62.PMID:6180969DOI:10.1007/BF02796473.
Cultured endothelial cells derived from bovine calf pulmonary artery were subjected to a variety of fixatives and stained with 1% Alcian Blue 8GX at pH 2.59 to 3.26. Within this range of pH, interphase nuclei and especially mitotic figures were (a) strongly stained in cells fixed with 10% formalin (phosphate buffered or unbuffered) or 2.5% buffered glutaraldehyde, (b) weakly stained or unstained in cells fixed in formaldehyde containing divalent cations, and (c) unstained in cells fixed in acetic acid-containing fluids. However, optimal nuclear staining with Alcian blue under the conditions of this study was judged to be achieved after fixation with neutral phosphate buffered 10% formalin. Endothelial cell cytoplasm exhibited a similar fixative-dependent staining. At pH 2.59 the cytoplasm of interphase cells fixed in formaldehyde (containing no divalent cations) or glutaraldehyde remained unstained; however, at higher pH cytoplasmic staining did occur and it increased as pH increased. In contrast, when these latter fixatives were employed the cytoplasm of mitotic cells stained at all pH levels tested. In cultured endothelial cells after appropriate fixation, 1% Alcian Blue 8GX (pH 2.59) was found to possess the ability to stain nuclei with a selectivity and intensity that compared favorably to those of the Feulgen reaction of Heidenhain iron hematoxylin but without the latters' length and complexity. Therefore, this procedure may provide a rapid, simple, and selective method for visualizing interphase nuclei or mitotic figures, or both in the majority of cultured cells.
Measurement of tissue disulfide groups by Alcian Blue 8GX and atomic absorption spectrophotometry
J Histochem Cytochem 1982 Dec;30(12):1311-2.PMID:6185563DOI:10.1177/30.12.6185563.
The primary purpose of this study was to quantify tissue disulfides with Alcian Blue 8GX using atomic absorption spectrophotometry (ASS) to measure the copper moiety of the Alcian blue. The secondary purpose was to determine whether aqueous or alcoholic formalin was a better fixative for this procedure. Sections of mouse renal cortex fixed in 10% neutral formalin (NBF) or in ethanol/formalin/acetic acid (AFA) were stained in 0.5% Alcian Blue 8GX in 0.8 M MgCl2 with prior thiosulfation (+Th) or without prior thiosulfation (-Th). The relative contents in the tissues of the copper of Alcian blue, measured by AAS, may be summarized as follows: -Th NBF, 1.15 micrograms Cu/mg tissue; -Th AFA, 1.35 micrograms Cu/mg tissue; +Th NBF, 1.99 micrograms Cu/mg tissue; +Th AFA, 3.25 micrograms Cu/mg tissue. These data indicate that 1) disulfide may be measured by this procedure, and 2) AFA is the preferred fixative for measurement of both disulfides and native sulfates.