Navigation Links
Fluorescence assay for determination of protease activity

In research and industrial work it is quite often necessary to test whether samples or solutions are free of protease activity. Using commercially available kits e.g. from Molecular Probes Inc., protease activity can be measured with the new fluorometer from PerkinElmer Life Sciences, VICTOR.

VICTOR is a new multitechnology and multilabel counter from Wallac that can measure fluorescent, photometric and luminescent labels as well as DELFIA research assays in microtitration plates. It is an epifluorometer that allows the use of both clear and coloured plates depending on the assay requirements.

DESCRIPTION OF ASSAY

In the reaction protease digests a highly quenched BODIPY FL-casein substrate releasing fluorescent peptides. The trypsin used as standard was diluted in 10 mmol/l Tris-HCl buffer, pH 7.8 of which 100 μl was added to the well.

After addition of 100 μl substrate, 10 μg/ml, diluted in 10 mmol/l Tris HCl buffer, pH 7.8, the plate was incubated overnight at room temperature and then measured.

DETECTION

The detection was done using the factory preset settings for fluorescein label. Excitation was at 485 nm and emission was measured at 535 nm.

RESULTS

Measurement with the new VICTOR fluorometer gives results comparable to the data given by the kit manufacturer.

BENEFITS

Fast and simple sample preparation.

PRODUCTS AVAILABLE

1420-011 VICTOR2 multilabel counter

1420-012 VICTOR2 multilabel counter with TR-fluorescence option

1420-014 VICTOR2 basic with stacker and robotic loading system

1420-015 VICTOR2 with TRF with stacker and robotic loading sy stem

1420-041 VICTOR2V with stacker

OTHER PRODUCTS NEEDED

Trypsin enzyme from bovine pancreas, activity 10700 units/mg (Sigma T-8253)

EnzCheck Protease Assay kit from Molecular Probes Inc. (E-6638) Labsystems black Combiplate (cat.no. 95029490)

REFERENCES

Insert of the EnzCheck Protease Assay Kit from Molecular Probes Inc. (E-6638).


'"/>

Source:


Page: All 1 2

Related biology technology :

1. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
2. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
3. Visualizing siRNA in Mammalian Cells: Fluorescence Analysis of the RNAi Effect
4. Identification of cells in S phase using the Cell Proliferation Fluorescence Kit and IN Cell Analyzer 1000
5. Minimizing Photobleaching of Blue Fluorescent Protein (BFP) using the Varian Cary Eclipse Fluorescence Spectrophotometer
6. Receptor Binding Assays using Fluorescence Polarization Detection: the [FP]2TM Characterized Tracers
7. TKXtra DataFile: Homogenous tyrosine kinase detection using High Efficiency Fluorescence Polarization
8. cAMP DataFile: Homogenous Cyclic AMP detection using High Efficiency Fluorescence Polarization
9. LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43)
10. FLIPR Calcium 3 Assay Kit - An Optimal No-Wash Fluorescence Calcium Indicator Reagent for Non-Optimal Calcium Responses
11. Enzyme Immunoassay for Studying Intracellular Levels of cAMP
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/4/2016)... EDISON, N.J. , Feb. 4, 2016 ... company focused on the development and commercialization of targeted ... BIO CEO & Investor Conference 2016, to be held ... and Source Capital Group,s 2016 Disruptive Growth & Healthcare ... on February 10-11, 2016. James Sapirstein ...
(Date:2/4/2016)... , February 4, 2016 ... Laboratories (ABL), Inc. --> Strasbourg, France ... --> PharmaVentures is pleased to announce that it ... its biopharmaceutical manufacturing unit in Strasbourg, France ... --> --> Transgene (Euronext: ...
(Date:2/3/2016)... , Feb. 3, 2016  With the ... patent cliff that is underway, therapies such as ... for a whole host of indications are in ... aid in the development and production of these ... poor quality and high costs, novel approaches and ...
(Date:2/3/2016)... ... 03, 2016 , ... Aerocom, a world-leading supplier of pneumatic ... American healthcare market. , Aerocom Healthcare, LLC will be based in Denver, Colo., ... new pneumatic tube systems or expand existing systems within the U.S. and Canada. ...
Breaking Biology Technology:
(Date:1/7/2016)... -- Various factors have contributed to the rise ... and biosimilars. Some of these factors include the ... for cost-effective alternatives, growing burden of chronic diseases, ... of their corresponding patented biologic drugs, and are ... efficacy. The global biosimilars market is estimated to ...
(Date:1/6/2016)... Jan. 6, 2016  Varam Capital, a provider of ... partner to deliver advanced authentication solutions to their clients. ... the poor. A loan of a few thousand rupees ... them the ability to purchase livestock or equipment for ... or stock for a local store. ...
(Date:1/6/2016)... --  A&D Medical , a global leader in connected ... unveiling a new line of personal mobile health devices ... – at CES 2016, with the introduction of its LifeSource ... With ... arm wireless blood pressure monitors represent the launch of ...
Breaking Biology News(10 mins):