Sample Files

AttoPhos

November 21, 2012

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Related Products: Gen5 for Detection

 

Gen5 version required: 1.0 or higher


Basis for the Assay:
 

Enzyme linked immunosorbent assays (ELISA) have been used to quantitate a wide range of compounds and pathogens for almost 40 years. One of the most commonly used enzyme conjugates is Alkaline Phosphatase (AP). (2’-[2-benzothiazoyl]-6’-hydroxybenzothiazole phosphate (BBTP) serves as a substrate for AP. Unreacted BBTP is weakly fluorescent, but in the presence of alkaline phosphatase it is converted to a highly fluorescent product, BBT, which has excitation and emission maxima of 435 nm and 575 nm respectively. ELISA assays that employ an alkaline phosphatase conjugate can be used with AttoPhos® substrate system.

The protocol calls for a fluorescence measurement using a 440/30 excitation filter and a 560/40 emission filter.

The experiment data file contains data from several unknown alkaline phosphatase samples using AttoPhos as the substrate.

Plate Configuration:

Plate configuration assumes that samples will be run in duplicate. The plate includes seven AP standards ranging from 0 to 0.05 DEA Units/well, and two blank wells. Thus, 40 samples can be run on a microplate in duplicate, in addition to the standard curve.

Report:

The report is configured to provide the (1) raw fluorescence, (2) blanked fluorescence, (3) calibration curve, (4) curve fitting results, and (5) unknown sample concentrations.

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