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A Sensitive, Continuously Recording Fluorogenic Assay for Calpain
- Source :
- Biochemical and Biophysical Research Communications. 248:293-296
- Publication Year :
- 1998
- Publisher :
- Elsevier BV, 1998.
-
Abstract
- We have developed a sensitive and continuously recording fluorogenic assay for the thiol protease calpain. This assay uses the dipeptide substrate Suc-Leu-Tyr-4-Methoxy-2-Naphthylamine (Suc-LY-MNA) in Tris buffer (pH 7.5) in the presence of 0.1% CHAPS. The assay is linear over a wide range of enzyme concentration and is capable of detecting 10 picomolar calpain making it more sensitive than any previously published method. Moreover, this assay gives a rate that is linear for over ten minutes making it useful for mechanistic studies of inhibitors. This assay can be easily adapted to a 96-well plate format facilitating the large scale screening of inhibitors.
- Subjects :
- Tris
Erythrocytes
Detergents
Biophysics
Naphthalenes
Sensitivity and Specificity
Biochemistry
Substrate Specificity
chemistry.chemical_compound
Chaps
Humans
Fluorometry
Enzyme Inhibitors
Molecular Biology
chemistry.chemical_classification
Chromatography
Dipeptide
biology
Calpain
Substrate (chemistry)
Cholic Acids
Dipeptides
Cell Biology
Recombinant Proteins
Kinetics
Enzyme
chemistry
biology.protein
Thiol Protease
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 248
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....bebf929bc5227ab9999d9b9f55974ca7