Biotin Immunoaffinity HPLC Column

The Biotin Immunoaffinity HPLC Column is For Research Use Only

Size: 3ml, 10 columns

Assay Principle

Many methods of biotin determination based on HPLC-UV (High Performance Liquid Chromatography Ultraviolet) detection may show either low sensitivity or low selectivity. This depends on the dilution factor of the matrices and if problematic matrices are applied.  This method of content determination of Biotin combines the high selectivity of affinity columns with its potential to concentrate elute and use high sensitivity biotin detection by post-column labeling with fluorescein- streptavidin conjugate.

Sample Preparation:
Biotin samples are to be extracted and analyzed with the method of Bachas et al. [N.G. Hentz, L.G. Bachas Methods Enzymol. 1997; 279:275-86], e.g. vitamin tablets, liquid vitamin preparations, cell culture extracts. Example: 25g vitamin containing tablets are dissolved in 100ml  PBS. The resulting extract may be filtered through a 0.45µm membrane filter.
Enrichment Step IAC:
4ml extract (containing the quantity of Biotin from a 1g sample of above-mentioned sample preparation is followed) is diluted with a total volume of 20ml PBS and then applied in a reservoir on top of the BioTeZ-Immunoaffinity Column. The optimal flow rate through the gel is between 1 to 3ml/min.

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Product Developed and Manufactured in Germany by BioTeZ Berlin-Buch GmbH