Sulfo-NHS-Biotin Kit
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
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.
• Amine-reaction—Reacting with primary amines (-NH2), such as lysine side-chains, or the amino-termini of polypeptides.
• Labeling antibody—This kit can label antibodies to facilitate immobilization, purification or detection.
• Labeling protein—This kit can label proteins to facilitate immobilization, purification or detection.
• Labeling Cell surface molecule—This kit can label the cell surface proteins because the negatively charged reagent does not permeate cell membranes.
• Irreversible-Permanent amide bonds are formed; Spacer arm cannot be cleaved.
• Very short arm—Spacer arm (total length added to target) is 13.5 angstroms; it consists of the native biotin valeric acid group only.
• 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.
| 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. | |||
| Label or Dye: | Biotin | Product Size: | 10 reactions |
| Labeling Scale: | 1-10 mg | Labeling Target: | Proteins (General), Antibodies (General) |
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 |
