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, 3-(4-carboxybenzoyl) quinoline -2- carboxaldehyde (CBQCA) reacts with primary amines of proteins to form fluorescent moieties. This compound has the added advantage of functioning well in the presence of lipids, which normally interfere with protein determinations. The fluorescent intensity is directly proportional to the total protein concentration present in the sample.
The protocol calls for a fluorescence measurement using a 460/40 excitation filter and a 560/40 emission filter.
The experiment data file contains all of the fluorescence determinations for a CBQCA total protein determination using BSA as the protein.
Plate configuration assumes that samples will be run in duplicate. Included is a standard curve of protein (0 to 1000 ng/well) as defined by Invitrogen’s CBQCA detection kit. The plate includes 7 standard concentrations, but no blank or controls wells. Thus, 41 samples can be run on a microplate in duplicate, in addition to the standard curve.
The report is configured to provide the (1) raw fluorescence, (2) standard curve, and (3) calculated catalase concentrations and statistics of unknown samples.