Sample Files

OPA Protein Quantitation

November 27, 2012

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Related Products: Gen5 for Detection



Gen5 version required: 1.0 or higher


Basis for the Assay:


Quantitation of total protein content is a measurement common to many applications in basic science and clinical research. The compound, σ-phthaldialdehyde (OPA) in conjunction with reduced sulfhydryl groups reacts with primary amines to form fluorescent moieties. This compound reacts very rapidly, but is much more water-soluble than fluorescamine. The fluorescence intensity is directly proportional to the total protein concentration present in the sample.

The protocol calls for a fluorescence measurement using a 360/40 excitation filter and a 460/40 emission filter.

The experiment data file contains all of the fluorescence determinations for an OPA total protein determination using BSA as the protein.

Plate Configuration:

Plate configuration assumes that samples will be run in duplicate. Included is a standard curve of protein (0 to 1000 μg/ml). The plate includes 8 standard concentrations, but has no blanks or controls 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) standard curve, and (3) calculated protein concentrations and statistics of unknown samples.

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