Related Experiment Videos
A sensitive colorimetric assay for human urinary kallikrein
Clinica Chimica Acta; International Journal of Clinical Chemistry
|January 31, 1980
Summary
A new substrate, L-Prolyl-L-phenylalanyl-L-arginine-alpha-naphthylester (Pro-Phe-Arg-NE), enables highly sensitive detection of human urinary kallikrein. This method allows kallikrein determination in small urine samples using a colorimetric assay.
Area of Science:
- Biochemistry
- Enzymology
Background:
- Kallikreins are serine proteases involved in various physiological processes.
- Accurate and sensitive assays for kallikrein activity are crucial for research and diagnostics.
Purpose of the Study:
- To synthesize a novel substrate, L-Prolyl-L-phenylalanyl-L-arginine-alpha-naphthylester (Pro-Phe-Arg-NE).
- To develop a highly sensitive colorimetric assay for human urinary kallikrein using the new substrate.
Main Methods:
- Synthesis of Pro-Phe-Arg-NE.
- Development of a colorimetric assay measuring alpha-naphthol release.
- Application of the assay for human urinary kallikrein detection.
- Preparation of kallikrein zymograms using polyacrylamide disc gel isoelectric focusing.
Main Results:
- The Pro-Phe-Arg-NE assay demonstrated high sensitivity, with a minimum detectable concentration of human urinary kallikrein at approximately 0.001 KU.
- The assay requires only a 25-microliter sample of human urine.
- Zymography revealed six distinct bands of human urinary kallikrein activity.
Conclusions:
- Pro-Phe-Arg-NE is an effective substrate for a sensitive kallikrein assay.
- The developed assay offers a precise method for quantifying human urinary kallikrein.
- This assay facilitates the study of kallikrein in biological samples.