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"We only use secondary antibodies from Jackson ImmunoResearch whenever possible! Customer service is also excellent and the price point for their products is very competitive."
Elizabeth Soberg, University of Washington
Biotin is a small molecule which non-covalently binds to avidin and streptavidin with very high affinity. The affinity of the interaction makes biotin an excellent conjugate for detection when used in immunohistochemistry techniques. Biotin conjugates can be employed in signal enhancement techniques.
Biotin-SP™ is our trade name for biotin with a 6-atom spacer positioned between biotin and the protein to which it is conjugated. When biotin-SP-conjugated antibodies are used in enzyme immunoassays, there is an increase in sensitivity compared to biotin-conjugated antibodies without the spacer. This is especially notable when Biotin-SP conjugated antibodies are used with alkaline phosphatase-conjugated streptavidin. Apparently, the long spacer extends the biotin moiety away from the antibody surface, making it more accessible to binding sites on streptavidin.
Biotinylated antibodies require additional reagents for visualization. We offer streptavidin and Mouse Anti-Biotin conjugated to fluorophores and enzymes.
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Labeled streptavidin-biotin (LSAB) method for signal amplification.
A. Label antigen with primary antibody.
B. Incubate with biotinylated secondary antibody
C. Incubate with conjugated streptavidin.
Anti-GFP immunohistochemical staining using Biotin-SP-conjugated Donkey Anti-Rabbit IgG (H+L) secondary antibody followed by Peroxidase-conjugated Streptavidin: (A) At 4 weeks post-transplantation, no GFP signal could be detected in the in vivo specimens. (B) GFP expressing cells were visualized by brown staining in the positive control; signal was visualized using the chromogenic substrate 3,3’ diaminobenzidine (DAB) substrate. Nuclei were counter-stained with hematoxylin.