| 包装: | 10 reactions |
| 市场价: | 2160元 |
The Sulfo-NHS-Biotin Kit contains reagents sufficient for 10 biotin labeling reactions. It is a water-soluble biotin reagent for labeling protein, antibody or other molecules with amidogen (NH2-). The biotin in the kit is sufficient and has been activated for direct use. The whole procedure is simple and quick.
| Sufficient For: 10 labeling reactions, each with 1 to 10 mg of antibody | |||
| Streptavidin | 10 mg | Sulfo-NHS-Biotin | 25 mg |
| HABA Solution | 1 mL | PBS Pack (makes 500 mL) | 1 pack |
| Sephadex G-25 in PD-10 Desalting Columns | 8.3 mL, 10 columns | ||
| Store biotin and streptavidin reagents at -20°C. Store remaining kit components at 4°C. | |||
o Amine-reaction—Reacting with primary amines (-NH2), such as lysine side-chains, or the amino-termini of polypeptides.
o Labeling antibody—This kit can label antibodies to facilitate immobilization, purification or detection.
o Labeling protein—This kit can label proteins to facilitate immobilization, purification or detection.
o Labeling Cell surface molecule—This kit can label the cell surface proteins because the negatively charged reagent does not permeate cell membranes.
o Irreversible-Permanent amide bonds are formed; Spacer arm cannot be cleaved.
o Very short arm—Spacer arm (total length added to target) is 13.5 angstroms; it consists of the native biotin valeric acid group only.
o Solubility increased—Sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds.
Sulfo-NHS-Biotin (N-Hydroxysulfosuccinimidobiotin) is an amine-reactive biotinylation reagent. The most common amine-reactive biotinylation reagents can be divied into two basic types, N-hydroxysuccinimide (NHS) esters and carboxylates. Generally, NHS esters biotinylation reagent contains a reactive NHS ring structure that reacts with an amine on the carbonyl group of a protein or other molecules through nucleophilic attack subsequently forming a stable amide linkage and releasing the NHS group. Due to the insolubility of NHS-biotin in aqueous environments, sulfo-NHS-biotin, a water-soluble analog of NHS-biotin, has been developed by attaching a negatively charged sulfonate group on the NHS ring structure, which can be added directly to aqueous reactions without the need for organic solvent dissolution.
1）Q：货号K1001-K1006的生物素化试剂盒中，生物素产品与哪些基团反应进行生物素化？
A：该系列试剂盒中生物素试剂（如Sulfo-NHS-LC-Biotin）上的NHS酯与伯胺反应。在蛋白质和多肽中，伯胺基团一般存在于赖氨酸残基的氨基末端和ε胺（侧链）上。
2）Q：带有磺酸基（Sulfo-）的Biotin与不带的Biotin有什么不同？
A：磺酸基（Sulfo-）Biotin可增加水溶性，常用于细胞表面标记；non-sulfonated Biotin需用DMSO或DMF等有机溶剂溶解，可透过细胞膜，常用于标记细胞内带有伯胺（NH2-）的蛋白。
3）Q：K1001-K1006试剂盒中Biotin产品有什么不同？
A：第一，带磺酸基（Sulfo-）的Biotin与不带Sulfo-的Biotin膜渗透性、水溶性不同；第二，(Sulfo) -NHS-Biotin试剂是一种短链生物素化试剂，(Sulfo) -NHS-LC-Biotin试剂是一种中等长度的生物素化试剂， (Sulfo) -NHS-SS-Biotin是一种间隔臂包含可裂解的二硫键的生物素化试剂。
4）HABA分析常见问题
| 常见问题 | 可能原因 | 解决方案 |
| HABA分析时 ΔA500 ≤0 | 蛋白质样品没有生物素化或生物素化水平很低，因为蛋白质上可标记的官能团有限 | 使用替代化学方法（例如巯基反应性-）重复生物素化或使用更高摩尔比的生物素化试剂 |
| 试剂混合不充分 | 稀释前完全溶解并混合HABA/链霉亲和素溶液 | |
| 样品中有颗粒 | 过滤蛋白质样品以去除颗粒 | |
| 生物素化水平偏高 | 未反应的生物素未去除 | 分析前对样品进行透析或脱盐 |
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