Absorption: (in nm) 500
Unit Price (USD):
1 kit $120.75
Bulk Price/unit (when you buy 5 or more): $96.60
Description:
Allows the measurement of the molar ratio of biotin to protein in biotin-protein conjugates, biotin-DNA molar ratios in biotinylated DNA samples, or quantitation of biotin concentrations in solution. The kit can be used to determine biotin concentrations down to 1 uM concentrations. The assay can be performed in a microtiterplate format.
Note:
The Kit includes enough reagents for 100 assays using a microtiterplate format (96-well plate, 200uL assay volume) or 20 assays using a spectrophotometer (1mL assay volume). A detailed protocol for use is also included.
Application:
This assay employs the sensitive chromogenic reagent 4'-hydroxyazobenzene-2-carboxylic acid (HABA) that exhibits a UV spectral absorbance at 500 nm when bound to avidin. Because biotin easily displaces HABA from HABA/Avidin complexes, the resulting decrease in absorption at 500 nm can be used to sensitively measure biotin concentrations.
References:
Batchelor RH, Sarkez A, Cox WG, Johnson I (2007) "Fluorometric assay for quantitation of biotin covalently attached to proteins and nucleic acids." Biotechniques 43(4): 503-7.
Ikariyama Y, Aizawa M (1988) "Bioaffinity sensors." Methods Enzymol. 137: 111-24.
Livnah O, Bayer EA, Wilchek M, Sussman JL (1993) "The structure of the complex between avidin and the dye, 2-(4'-hydroxyazobenzene) benzoic acid (HABA)." FEBS Lett. 328(1-2): 165-8.
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