Biotin (Long Arm) Maleimide is designed for biotinylating proteins, nucleic acids or other molecules containing one or more thiol groups. The presence of the 6-aminohexanoate spacer arm between the maleimide group and biotin reduces the possibility of steric hinderance. Molecules to be labeled with Biotin (Long Arm) Maleimide require free thiol groups. In the case of proteins, Biotin (Long Arm) Maleimide will react with exposed cysteine residues. Alternatively, free thiols may be generated by reducing disulfide bonds or by modifying other reactive groups such as priamry amines with componds like Traut’s reagent (2-iminothiolane). For labeling nucleic acids, thiols can be introduced into DNA, RNA, or oligonucleotides using the FastTag, 5’EndTag or 3’ EndTag nucleic acid labeling systems. Because of the extraordinary affinity of avidin and streptavidin to biotin and the many biotin-avidin/streptavidin systems available, this label is ideal for a variety of applications including in situ hybridization, blotting, and affinity binding. Biotin (Long Arm) Maleimide reacts readily under mild acid or neutral pH conditions in aqueous solution with free sulfhydryl groups, such as those of cysteine residues or sulfhydryls introduced chemically into proteins or other macromolecules. The spacer arm in this derivative insures that the biotin is fully accessible to avidin or streptavidin detection systems.
Target for Labeling:DNA, Oligonucleotides, PNA, Protein, RNA
Required Reactive Group:Thiol Group
Tag/Group Incorporated:Biotin
Molecular Formula:C22H33N5O5S
Applications:Suitable for use in ELISA. Other applications not tested.
Recommended Dilutions:Optimal dilutions to be determined by the researcher.
Reconstitution of B1750-51:FastTag® labeling of nucleic acids: Reconstitute with 500ul anhydrous DMF. NOTE: It is not recommended to store the solution.
5’ EndTag™ or 3’ EndTag™ labeling of nucleic acids: Reconstitute with 312ul anhydrous DMF. NOTE: It is not recommended to store the solution.
Follow the labeling procedure included with the 5’ EndTag™ or 3’ EndTag™ systems.
Protein Labeling Procedure:1. Dissolve the protein to be labeled in 100mM PBS, pH 7.0, to a concentration of 5mg/ml
2. Dissolve a slight excess of B1750-51 in DMF at a concentration of 20mg/ml
3. Add 25ul/ml of B1750-51 of protein solution
4. Icubate at RT for 3 hours with occasional stirring
5. Separate the unreacted material from the protein by gel filtration or dialysis.
Storage and Stability:Lyophilized product is stable for 6 months after receipt at -20°C. Reconstitute with sterile DMF. It is not recommended to store reconstituted solution.
