Dojindo,Biotin Sulfo-OSu/10/B319,Biotin-(AC5)2-OSu用作 Biotin Labeling Kit-NH2 中的生物素标记剂

抗生物素蛋白-生物素系统在免疫学和组织化学中有许多应用。抗生物素蛋白和生物素之间的相互作用非常强，解离常数约为 10-15 M。生物素通常添加到一抗或二抗中，例如抗 IgG 和抗 IgM。在用生物素标记的抗体制备抗原-抗体复合物后，使用酶或荧光素标记的抗生物素蛋白或链霉抗生物素蛋白对抗原进行比色或荧光检测。琥珀酰亚胺酯生物素在 pH 7-9 下与伯胺和仲胺（例如氨基酸和蛋白质）反应。琥珀酰亚胺酯与游离胺基反应生成稳定的酰胺键。琥珀酰亚胺基生物素试剂必须溶解在 DMSO、DMF 或酒精中。用 DMSO 制备的储备溶液在 -20ºC 下可稳定数月。磺基琥珀酰亚胺生物素试剂可溶于水，因此无需使用 DMF 或 DMSO 等有机溶剂。使用具有较长间隔物的生物素（如 Biotin-(AC5)2-OSu 或 Biotin-(AC5)2-Sulfo-OSu）制备的 IgG 具有更好的信噪比。较长的间隔使链霉亲和素或抗生物素 IgG 能够在没有结构抑制的情况下识别生物素。因此，Biotin-(AC5)2-OSu 被用作 Biotin Labeling Kit-NH2 中的生物素标记剂。

Labeling Procedure for IgG1. Prepare 10 mM of the biotin labeling reagent using DMSO.2. Prepare 100 μl of 1 mg per ml IgG buffer solution (pH 7.5-8.5) that does not contain any large molecules with amine compounds.3. Add 1-5ml biotin labeling reagent DMSO solution to the IgG buffer solution and incubate at 37ºC for 1 hour.4. Remove excess biotin labeling reagent using gel filtration or dialysis.5. Prepare solutions for further experiment using an appropriate buffer such as PBST (0.05% Tween 20/PBS).

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