Biotin is a small biomolecule important for many cellular processes. Most importantly for biotechnology applications, biotin is ammenable to conjugation to proteins for use in biochemical assays. Biotin has a very strong affinity for avidin and streptavidin; an attraction that is the strongest and most stable non-covalent interaction known. B-galactosidase (b-gal; b-gal) is an enzyme that hydrolyzes b-galactosides (such as lactose and various glycoproteins) into simple sugars. B-galactosidase is routinely used as a reporter in biochemical experiments as well as a marker for gene expression experiments in E. coli. B-galactosidase can be used to cleave the chemical X-gal that forms an intense blue product that can easily be quantified.
Source:
B-Galactosidase, expressed in E.coli.
Applications:
Suitable for use in ELISA, Western Blot and Immunohistochemistry. Other applications not tested.
Recommended Dilution:
ELISA: 1:8000-1:32,000
Western Blot: 1:500-1:2500
Immunohistochemistry: 1:200-1:1000
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C. For long-term storage, aliquot and store at 4 degrees C. Do not freeze. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.