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218693 Protease Substrate

Casein labeled with N-(resorufin-4-carbonyl)piperidine-4-carbonic acid.

Protease Substrate MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: Casein, Resorufin-Labeled

218693
  
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Overview

Replacement Information
Description
Overview

This product has been discontinued.





Casein labeled with N-(resorufin-4-carbonyl)piperidine-4-carbonic acid. Colorimetric or fluorometric substrate for sensitive detection of protease contamination in solution.
Catalogue Number218693
Brand Family Calbiochem®
SynonymsCasein, Resorufin-Labeled
References
Product Information
FormRed lyophilized solid
Applications
Biological Information
Physicochemical Information
Emission max.
Excitation max.
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Ambient Temperature Only
Toxicity Standard Handling
Storage -20°C
Do not freeze Ok to freeze
Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C) for long term storage or refrigerate (4°C) for short term storage. Stock solutions are stable for up to 3 days at 4°C and for up to 3 months at -20°C.
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
218693 0

Documentation

Protease Substrate SDS

Title

Safety Data Sheet (SDS) 

Protease Substrate Certificates of Analysis

TitleLot Number
218693
Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision03-June-2008 RFH
SynonymsCasein, Resorufin-Labeled
DescriptionCasein labeled with N-(resorufin-4-carbonyl)piperidine-4-carbonic acid. Colorimetric and fluorometric substrate for sensitive detection of protease contamination in solution.
FormRed lyophilized solid
Recommended reaction conditions
Assay for Determination of Proteolytic Activity Note: This protocol is provided only as a general guide. Researchers should standardize this assay for their own specific needs and should consult published literature. Introduction: Protease substrate is milk casein that is coupled with activated resorufin (N-(resorufin-4-carbonyl)piperidine-4-carbonic acid N-hydroxysuccinimide ester). It is a general protease substrate and is particularly useful for the detection of trace protease activity. Treatment of the substrate with protease results in the release of resorufin-labeled peptides that are not precipitated by trichloroacetic acid (TCA). The concentration of the labeled peptides in the supernatant is proportional to the proteolytic activity in the sample. The following protocol has been modified according to Twining, S.S. Reagents and Equipment: • Sample solution • Substrate solution: 0.4% protease substrate (casein, resorufin-labeled, Cat. No. 218693) in dH2O • Incubation buffer: 0.2 M Tris-HCl, pH 7.8, 0.02 M CaCl2 • Stop buffer: 5% Trichloroacetic acid (w/v) in dH2O • Assay buffer: 0.5 M Tris-HCl, pH 8.8 • Spectrophotometer capable of measuring absorbance at 574 nm (colorimetric) or fluorimeter capable of measuring excitation at 574 nm and emission at 584 nm (fluorometric) • Incubator capable of maintaining constant temperature at 37°C Protocol: • For each sample, mix the following: 50 µl substrate solution 50 µl incubation buffer 100 µl sample solution • Prepare a blank by mixing the following: 50 µl substrate solution 50 µl incubation buffer 100 µl dH2O • Incubate samples and blank at 37°C for the desired amount of time (15 min to overnight) Note: The incubation time will depend on the amount of protease present in the unknown sample(s). It may be necessary to perform a time-course study to determine the optimal incubation time for a given sample. Various concentrations of protease have yielded the following results: Concentration        Color Change Within 2 mg/ml                         ~1 min. 0.5 mg/ml                      ~5 min. 0.2 mg/ml                      ~10 min. • Stop the reactions by adding 480 µl of stop buffer (samples and blank) • Incubate samples and blank at 37°C for 10 min • Centrifuge for 5 min and transfer 400 µl of each supernatant to a cuvette • Add 600 µl of assay buffer, mix, and immediately read the absorbance of the sample against the blank at 574 nm, 15-25°C. The color will change from a bluish-violet to red. • Alternatively, the sensitivity may be improved by measuring the cleavage in a fluorimeter with an excitation wavelength of 574 nm and an emission wavelength of 584 nm
SolubilityH₂O (4 mg/ml)
Storage -20°C
Do Not Freeze Ok to freeze
Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C) for long term storage or refrigerate (4°C) for short term storage. Stock solutions are stable for up to 3 days at 4°C and for up to 3 months at -20°C.
Toxicity Standard Handling