标签：抗生物素蛋白-生物素系统

Dojindo,Biotin-AC5 Sulfo-OSu/10/B320,抗生物素蛋白-生物素系统

Product Description of Amine-Reactive Biotins
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).

1. J. Wormmeester, F. Stiekema and C. Groot, Immunoselective Cell Separation, Methods Enzymol., 1990, 184, 314.2. J. J. Leary, D. J. Brigati and D. C. Ward, Rapid and Sensitive Colorimetric Method for Visualizing Biotin-labeled DNA Probes Hybridized to DNA or RNA Immobilized on Nitrocellulose: Bio-blots, Proc. Batl. Acad. Sci. USA, 1983, 80, 4045.3. W. T. Lee and D. H. Conrad, The Murine Lymphocyte Receptor for IgE. II. Characterization of the Multivalent Nature of the B Lymphocyte Receptor for IgE, J. Exp. Med., 1984, 159, 1790.4. D. R. Gretch, M. Suter and M. F. Stinski, The use of Biotinylated Monoclonal Antibodies and Streptavidin Affinity Chromatography to Isolate Herpesvirus Hydrophobic Proteins or Glycoproteins, Anal. Biochem., 1987, 163, 270 .5. M. Shimkus, J. Levy and T.Herman, A Chemically Cleavable Biotinylated Nucleotide: Usefulness in the Recovery of Protein-DNA Complexes from Avidin Affinity Columns, Proc. Natl. Acad. Sci. USA, 1985, 82, 2593.6. W. J. LaRochelle and S. C. Froehner, Immunochemical Detection of Proteins Biotinylated on Nitrocellulose Replicas, J. Immunol. Methods, 1986, 92, 65.7. P. S. Anjaneyulu and J. V. Staros, Reactions of N-hydroxysulfosuccinimide Active Esters, Int. J. Pep. Protein Res., 1987, 30, 117.8. H. M. Ingalls, C. M. Goodloe-Holland and E. J. Luna, Junctional Plasma Membrane Domains Isolated from Aggregating Dictyostelium Discoideum Amebae, Proc. Natl. Acad. Sci. USA, 1986, 83, 4779.9. J. L. Guesdon, T. Ternynck and S. Avrameas, The Use of Avidin-biotin Interaction in Immunoenzymatic Techniques, J. Histochem, Cytochem, 1979, 27, 1131.

Dojindo,Biotin-PEAC5-maleimide/10/B299,抗生物素蛋白-生物素系统

Product Description of Amine-Reactive Biotins
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。生物素通常添加到抗 IgG 和抗 IgM 等一抗或二抗中。在用生物素标记的抗体制备抗原-抗体复合物后，使用酶或荧光团标记的抗生物素蛋白或链霉抗生物素蛋白进行抗原的比色或荧光检测。马来酰亚胺生物素在 pH 7-7.5 下与硫醇化合物反应，例如具有巯基的蛋白质或肽。马来酰亚胺与巯基反应生成硫醚键。虽然其他马来酰亚胺生物素试剂必须溶解在 DMSO、DMF 或酒精中，但生物素-PE-马来酰亚胺可以在 pH 7.4 的 PBS 中溶解，以制备 2 mM 溶液，而无需使用有机溶剂。马来酰亚胺与巯基的反应性高于溴乙酰胺，因此马来酰亚胺生物素所需浓度远低于溴乙酰胺生物素。 Biotin-PE-maleimide 和 Biotin-PEAC5-maleimide 在 DMSO 中的储备溶液在 -20oC 下可稳定保存一年。

Structural Formula:

Labeling Procedure for Reduced IgG1. Prepare 10 mM of the biotin labeling reagent using DMSO.2. Prepare 100 μl of 1 mg per ml reduced IgG/ml buffer solution which does not contain any large molecules with SH groups. Reduced IgG can be prepared by TCEP (tricarboxyethylphosphine), DTT or 2-mercaptoethylamine.3. Add 1-5 μl of biotin labeling reagent DMSO solution to the IgG buffer solution and incubate at 37 oC for 1 hour.4. Remove excess biotin labeling reagent using a gel column or a Filtration tube.5. Prepare solutions for further experiment using an appropriate buffer such as PBST (0.05% Tween 20/PBS).

E. Muneyuki, et al., Biophys. J., 92, 1806 (2007).

Dojindo,Biotin OSu/10/B304,抗生物素蛋白-生物素系统

Product Description of Amine-Reactive Biotins
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。

Reaction Scheme

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).

1. P. Kongtawelert and P. Ghosh, A New Sandwich-ELISA Method for the Determination of Keratan Sulphate Peptides in Biological Fluids Employing a Monoclonal Antibody and Labelled Avidin Biotin Technique., Clin. Chem. Acta,, 1990, 195, 17 .2. G. Paganelli, S. Pervez, A. G. Siccardi, G. Rowlinson, G. Deleide, F. Chiolerio, M. Malcovati, G. A. Scassllati and A. A. Epenetos, Intraperitoneal Radio-localization of Tumors Pre-targeted by Biotinylated Monoclonal Antibodies, Int. J. Cancer, 1990, 45, 1184.3. C. Wagener, U. Kruger and J. E. Shively, Selective Precipitation of Biotin-labeled Antigens or Monoclonal Antibodies by Avidin for Determining Epitope Specificities and Affinities in Solution-phase Assays, Methods Enzymol., 1990, 184, 518 .4. D. M. Boorsma, J. Van Bommel and E. M. Vander Raaij-Helmer, Simultaneous Immunoenzyme Double Labelling Using Two Different Enzymes Linked Directly to Monoclonal Antibodies or with Biotin-avidin, J. Microscopy, 1986, 143, 197.5. A. Komura, T. Tokuhisa, T. Nakagawa, A. Sasase, M. chihashi, S. Ferrone and Y.Mishima, Specific Killing of Human Melanoma Cells with an Efficient 10B-compound on Monoclonal Antibodies, Pigment Cell Res., 1989, 2, 259.6. R. Rappuoli, P. Leoncini, P. Tarli and P. Neri, Competitive Enzyme Immunoassay for Human Chorionic Somatomammotropin Using the Avidin-biotin System, Anal. Biochem., 1981, 118, 168.

Dojindo,Biotin-(AC5)2-OSu/10/B306,抗生物素蛋白-生物素系统

Product Description of Amine-Reactive Biotins
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).

1. P. Kongtawelert and P. Ghosh, A New Sandwich-ELISA Method for the Determination of Keratan Sulphate Peptides in Biological Fluids Employing a Monoclonal Antibody and Labelled Avidin Biotin Technique., Clin. Chem. Acta,, 1990, 195, 17 .2. G. Paganelli, S. Pervez, A. G. Siccardi, G. Rowlinson, G. Deleide, F. Chiolerio, M. Malcovati, G. A. Scassllati and A. A. Epenetos, Intraperitoneal Radio-localization of Tumors Pre-targeted by Biotinylated Monoclonal Antibodies, Int. J. Cancer, 1990, 45, 1184.3. C. Wagener, U. Kruger and J. E. Shively, Selective Precipitation of Biotin-labeled Antigens or Monoclonal Antibodies by Avidin for Determining Epitope Specificities and Affinities in Solution-phase Assays, Methods Enzymol., 1990, 184, 518 .4. D. M. Boorsma, J. Van Bommel and E. M. Vander Raaij-Helmer, Simultaneous Immunoenzyme Double Labelling Using Two Different Enzymes Linked Directly to Monoclonal Antibodies or with Biotin-avidin, J. Microscopy, 1986, 143, 197.5. A. Komura, T. Tokuhisa, T. Nakagawa, A. Sasase, M. chihashi, S. Ferrone and Y.Mishima, Specific Killing of Human Melanoma Cells with an Efficient 10B-compound on Monoclonal Antibodies, Pigment Cell Res., 1989, 2, 259.6. R. Rappuoli, P. Leoncini, P. Tarli and P. Neri, Competitive Enzyme Immunoassay for Human Chorionic Somatomammotropin Using the Avidin-biotin System, Anal. Biochem., 1981, 118, 168.