Search Results of “Enzyme Assay”:
Proteolytic Enzymes for Peptide Production
Author(s): Patricia J. Sweeney, John M. Walker
Pub. Date: May-22-1993; DOI:10.1385/0-89603-234-5:277
Summary: . The amino acid sequence is known (7). 2.1.4 pH Optimum The enzyme has a pH optimum between 7.5 and 8.5 (2). 2.1.5 Assay The assay is based on measuring the esterolytic activity of the enzyme...
Abstract | Full Text | PDF (1749K)
Herpesvirus Protease Assays
Author(s): Peter Ertl, Linda Russell, Jane Angier
Pub. Date: Sept-14-1999; DOI:10.1385/1-59259-245-7:171
Summary: for their ability to inhibit the protease enzyme. A number of assay formats have been used to examine the effect of herpesvirus protease inhibitors. This chapter describes four of these assays...
Abstract | Full Text | PDF (287K)
Characterization of Enzyme Activity, Protein Content, and Thiol Groups in Immobilized Enzymes
Author(s): Gordon F. Bickerstaff
Pub. Date: Dec-01-1996; DOI:10.1385/0-89603-386-4:253
Summary: that the normal procedures used for assay of the soluble enzyme activity, protein content, and so on, must be redesigned to accomodate the presence of the support material. In general, problems are likely...
Abstract | Full Text | PDF (554K)
Reporter Enzyme Assays
Author(s): Karen Pardy
Pub. Date: May-21-1993; DOI:10.1385/0-89603-245-0:419
Summary: the advantage that it has no mammalian counterpart. The enzyme catalyzes the following reaction: Chloramphenicol + acetyl CoA → 3-acetylchloramphenicol + CoA The CAT assay was originally...
Abstract | Full Text | PDF (298K)
Aminopeptidases: Aminopeptidase M (EC 3.4.11.2), Pyroglutamate Aminopeptidase (EC 3.4.19.3), and Prolidase (EC 3.4.13.9)
Author(s): Patricia J. Sweeney, John M. Walker
Pub. Date: May-22-1993; DOI:10.1385/0-89603-234-5:319
Summary: Optimum The enzyme is active at pH 7-9, but is normally used at pH 8.0 (22). 2.1.5 Assay The assay is based on the hydrolysis of L-pyroglutamic acid β-naph-thylamide. The enzyme buffer used is prepared...
Abstract | Full Text | PDF (1153K)
Improvement, Modification, Adaptation, Troubleshooting, and Development of New Methods
Pub. Date: Feb-08-1993; DOI:10.1007/978-1-60327-407-4_8
Summary: . This is important in any assay, but is usually of particular importance in an enzyme assay. Reagent pH Ordinarily, it is desirable to measure an enzyme at its optimal pH. This is not just because it gives maximal...
Abstract | Full Text | PDF (1545K)
Immobilization of Enzymes by Covalent Attachment
Author(s): Scott J. Novick, J. David Rozzell
Pub. Date: Jan-19-2005; DOI:10.1385/1-59259-846-3:247
Summary: enzyme that may be entrapped in the pores of the particles or loosely bound through noncovalent interactions. 1.5.1 Activity Assay There are two basic methods to measure activity-batch and continuous...
Abstract | Full Text | PDF (253K)
Acetate Kinase From Methanosarcina thermophila, a Key Enzyme for Methanogenesis
Author(s): Prabha Iyer, James G. Ferry
Pub. Date: Jan-19-2005; DOI:10.1385/1-59259-846-3:239
Summary: is a method for routine assay of the enzyme, and enzymelinked assay procedures for obtaining kinetic constants in the forward and reverse reaction directions. Finally, a method is included...
Abstract | Full Text | PDF (111K)
Development of a Hepatitis C Virus RNA Helicase High Throughput Assay
Author(s): Ann D. Kwong, Christine Risano
Pub. Date: Sept-14-1999; DOI:10.1385/1-59259-245-7:97
Summary: , substrate, and enzyme concentration, and is prevented by the presence of an inhibitor. The assay format can utilize double-stranded DNA as well as RNA substrates, making the assay amenable to use...
Abstract | Full Text | PDF (490K)
Studies of Enzymes That Cause Resistance to Aminoglycosides Antibiotics
Author(s): Engin H. Serpersu, Can Özen, Edward Wright
Pub. Date: Dec-01-2007; DOI:10.1007/978-1-59745-246-5_20
Summary: is A discontinuous coupled enzyme assay based on the conversion of one molecule of the reaction product, inorganic pyrophosphate, to two molecules of inorganic phosphate was utilized to measure the steady...
Abstract | Full Text | PDF (374K)
Identifying and Characterizing HIV Protease Inhibitors
Author(s): Eric S. Furfine
Pub. Date: Sept-14-1999; DOI:10.1385/1-59259-245-7:313
Summary: outlines the analysis required to deal with these potent compounds. Like any enzyme assay, preparing this assay for routine use can be divided into three parts: (1) determination of an appropriate...
Abstract | Full Text | PDF (297K)
Mycobacterium tuberculosis β-Ketoacyl Acyl Carrier Protein Synthase III (mtFabH) Assay: Principles and Method
Author(s): Sarbjot Sachdeva, Kevin A. Reynolds
Pub. Date: Dec-01-2007; DOI:10.1007/978-1-59745-246-5_16
Summary: and structures like phospholipid biosynthesis, cell wall formation, etc. Herein we describe a new assay for the Mycobacterium tuberculosis FabH (mtFabH) enzyme involved in a key initiation step in the synthesis...
Abstract | Full Text | PDF (437K)
Adsorption of Lipase on Inorganic Supports
Author(s): José V. Sinisterra
Pub. Date: Dec-01-1996; DOI:10.1385/0-89603-386-4:327
Summary: °C. 3. Assay the residual enzyme activity by adding a volume of immobilized enzyme to 10 mL of substrate emulsion of olive oil at 37°C (see Note 5). 4. Incubate the immobilized enzyme and olive oil...
Abstract | Full Text | PDF (175K)
Immobilization-Stabilization of Enzymes by Multipoint Covalent Attachment on Supports Activated With Epoxy Groups
Author(s): Cesar Mateo, Olga Abian, Gloria Fernández-Lorente, Benevides C. Pessela, Valeria Grazu, Jose M. Guisan, Roberto Fernandez-Lafuente
Pub. Date: Mar-15-2006; DOI:10.1007/978-1-59745-053-9_4
Summary: stirring at 25°C. (see Note 6 ) 3. Periodic samples of the supernatant and suspension were taken for assay of enzyme activity. Supernatant was achieved either by using pipet filter or by centrifugation...
Abstract | Full Text | PDF (175K)
Assay of N 8-Acetylspermidine Deacetylase
Author(s): Jim Blankenship
Pub. Date: Oct-10-1997; DOI:10.1385/0-89603-448-8:77
Summary: Assay of N 8-Acetylspermidine Deacetylase N 8-Acetylspermidine deacetylase is a cytoplasmic enzyme found in a wide variety of tissues in higher organisms (1...
Abstract | Full Text | PDF (509K)
Colorimetric Assays for Screening Laccases
Author(s): Miguel Alcalde, Thomas Bulter
Pub. Date: May-16-2003; DOI:10.1385/1-59259-396-8:193
Summary: of PAH-activity is the amount of enzyme that produces 1 μmol of 9,10-anthraquinone/min under the described conditions. 3.2.2 Decolorization Assay (see Note 11 ) A second colorimetric assay based...
Abstract | Full Text | PDF (195K)
Immobilization and Stabilization of Proteins by Multipoint Covalent Attachment on Novel Amino-Epoxy-Sepabeads®
Author(s): Cesar Mateo, Benevides C. Pessela, Valeria Grazu, Fernando López-Gallego, Rodrigo Torres, Manuel Fuentes, Aurelio Hidalgo, Jose M. Palomo, Lorena Betancor, Gloria Fernández-Lorente, Claudia Ortiz, Olga Abian, Jose M. Guisan, Roberto Fernandez-Lafuente
Pub. Date: Mar-15-2006; DOI:10.1007/978-1-59745-053-9_14
Summary: 3 ). 3. Periodically, samples of supernatant and suspension were taken for assay of enzyme activity. Supernatant was achieved by using a tip filter or by centrifugation of the suspension (see Note 4...
Abstract | Full Text | PDF (217K)
Poly(Ethylene Glycol) Crosslinked to Albumin as a Support for Enzyme Immobilization
Author(s): Guy Fortier, Nicole Demers, Jacques Jean-Fran çois, Jean-Charles Gayet, Edith M. D’Urso
Pub. Date: Dec-01-1996; DOI:10.1385/0-89603-386-4:117
Summary: nm. 13. Protein assay: Add 1 mL of the working reagent to 0.1 mL of the sample. Incubate at 37°C for 30 min and read the absorbance at 562 nm. 14. The enzyme hydrogel can now be tested for its...
Abstract | Full Text | PDF (427K)
Overview of Antigen Detection Through Enzymatic Activity
Author(s): Gary L. Bratthauer
Pub. Date: Oct-17-1994; DOI:10.1385/0-89603285-X:155
Summary: . A glucose oxidase system can provide a sensitive and specific assay if other endogenous enzyme activity is a problem. 3 Chromogenic Substrates In addition to the many enzyme systems available...
Abstract | Full Text | PDF (833K)
Screening for Thermostability
Author(s): Patrick C. Cirino, Radu Georgescu
Pub. Date: May-16-2003; DOI:10.1385/1-59259-396-8:117
Summary: are described in other chapters of this volume. In this section, we describe an assay for thermostability in which enzyme activity is measured using a 96-well plate absorbance or fluorescence reader...
Abstract | Full Text | PDF (96K)
Design and Implementation of High Throughput Screening Assays
Author(s): Ricardo Macarrón, Robert P. Hertzberg
Pub. Date: Apr-18-2002; DOI:10.1385/1-59259-180-9:001
Summary: in the detection system. If the enzyme is not stable at low concentrations, or if the assay method does not respond linearly to product formation or substrate depletion, there could also be a lack of linearity...
Abstract | Full Text | PDF (679K)
Nonradioactive Trans-Sialidase Screening Assay
Author(s): Silke Schrader, Roland Schauer
Pub. Date: Sept-15-2006; DOI:10.1385/1-59745-167-3:93
Summary: exist for the detection of the activity of this enzyme. Mattos-Guaraldi et al. (26) developed a peanut lectin hemagglutination assay for testing the TS from C. diphtheriae. An enzyme-linked immunoassay...
Abstract | Full Text | PDF (593K)
Immobilization of Enzymes on Electrodes
Author(s): Gilvanda Silva Nunes, Jean-Louis Marty
Pub. Date: Mar-15-2006; DOI:10.1007/978-1-59745-053-9_21
Summary: Immobilization of Enzymes on Electrodes This chapter focuses on the four main immobilization techniques used in the development of enzyme electrodes: adsorption, entrapment, covalent coupling...
Abstract | Full Text | PDF (189K)
Hyaluronidase Activity and Hyaluronidase Inhibitors: Assay Using a Microtiter-Based System
Author(s): Susan Stair Nawy, Antonei B. Csóka, Kazuhiro Mio, Robert Stern
Pub. Date: June-01-2001; DOI:10.1385/1-59259-209-0:383
Summary: hyaluronidase with a known activity, in the appropriate buffer, from 1.0 to 1 × 106 rTRU and assay 100 μL/well, the standard enzyme reaction volume, in triplicate (see Note 4 ). 3. Pipet unknown...
Abstract | Full Text | PDF (113K)
Screening Hybridoma Culture Supernatants Using ELISA
Author(s): Mark Page, Robin Thorpe
Pub. Date: Feb-15-2002; DOI:10.1385/1-59259-169-8:1117
Summary: Screening Hybridoma Culture Supernatants Using ELISA Enzyme-linked immunosorbent assay (ELISA) is a widely used method for the detection of antibody and is appropriate for use for screening...
Abstract | Full Text | PDF (79K)
O-Linked Chain Glycosyltransferases
Author(s): Inka Brockhausen
Pub. Date: Mar-01-2000; DOI:10.1385/1-59259-048-9:273
Summary: source of the desired enzyme under the conditions described for the standard transferase assay. 3. Low molecular weight compounds are isolated by gel filtration on Bio-Gel P4 or P2 columns, followed...
Abstract | Full Text | PDF (154K)
α-Galactosidase Assay in Fermented Soymilk Products
Author(s): Marisa S. Garro, Graciela Font de Valdez, Graciela Savoy de Giori
Pub. Date: July-15-2004; DOI:10.1385/1-59259-765-3:121
Summary: α-Galactosidase Assay in Fermented Soymilk Products 11.1 Introduction The enzyme alpha-galactosidase (α-gal) (EC. 3.2.1.22) hydrolyzes the α-1,6-galactosidic bonds present in melibiose...
Abstract | Full Text | PDF (400K)
Stabilization of Multimeric Enzymes Via Immobilization and Further Cross-Linking With Aldehyde-Dextran
Author(s): Cesar Mateo, Benevides C. Pessela, Manuel Fuentes, Rodrigo Torres, Lorena Betancor, Aurelio Hidalgo, Gloria Fernández-Lorente, Roberto Fernandez-Lafuente, Jose M. Guisan
Pub. Date: Mar-15-2006; DOI:10.1007/978-1-59745-053-9_12
Summary: . The assay was carried out in Novo buffer, pH (6.5) at 25°C. Values were reproduced in two separate experiments. All other conditions are detailed in Subheadings 2.. and 3. (●) Sepabeads-boronic-epoxy...
Abstract | Full Text | PDF (237K)
Hydrogen Peroxide Assay for Amine Oxidase Activity
Author(s): R. James Storer, Antonio Ferrante
Pub. Date: Oct-10-1997; DOI:10.1385/0-89603-448-8:81
Summary: dimer is more stable (19). Fig. 1. (A) Schematic for the coupled enzyme assay of spermine oxidation by amine oxidase (B) Oxidation of homovanillic acid by the peroxidase(HRPO)-H2O2 complex. The HRPO...
Abstract | Full Text | PDF (850K)
Enzyme Biosensors Based on Fluorometric Detection
Author(s): Ashutosh Sharma
Pub. Date: Apr-17-1998; DOI:10.1385/0-89603-410-0:187
Summary: enzymes, one of the most extensively exploited fluorescence-based enzyme assays relies on changes in the fluorescence of this cofactor. This assay method is based on the difference observed...
Abstract | Full Text | PDF (588K)
High-Throughput Screening of Human Cytochrome P450 Inhibitors Using Fluorometric Substrates: Methodology for 25 Enzyme/Substrate Pairs
Author(s): David M. Stresser
Pub. Date: Aug-11-2004; DOI:10.1385/1-59259-800-5:215
Summary: engineering (12), and plate-based CYP fluorometric assays (13). Since then, the assay has been optimized, validated, and expanded to include many other enzyme/substrate pairs (14–29), including CYP...
Abstract | Full Text | PDF (144K)
Proteinase K (EC 3.4.21.14)
Author(s): Patricia J. Sweeney, John M. Walker
Pub. Date: May-22-1993; DOI:10.1385/0-89603-234-5:305
Summary: hemoglobin, showed optimal activity in the pH range 7.5–12.0 (1). However, the enzyme is normally used in pH range 7.5–9.0 (1,26,27). 2.5 Assay The assay is based on the hydrolysis of N-acetyl...
Abstract | Full Text | PDF (761K)
A Systematic Approach for the Solubilization of the Integral Membrane Protein Lysophospholipid: Acyl-Coa Acyltransferase (LAT)
Author(s): Claus Kerkhoff, Volkhard Kaever
Pub. Date: May-29-2003; DOI:10.1385/1-59259-400-X:111
Summary: micelles were accessible to protein purification, and we are currently analyzing the protein composition of a LAT candidate protein. 2 Materials 2.1 LAT Standard Enzyme Assay 1. LAT assay buffer: 150 mM NaCl...
Abstract | Full Text | PDF (160K)
Enzyme-Linked Immunosorbent Assays
Author(s): David M. Kemeny
Pub. Date: Aug-28-1998; DOI:10.1385/0-89603-479-8:257
Summary: of antibody. In this assay competition is determined between (A) the sample antibody and a fixed amount of plate-bound antigen and enzyme-labeled antibody, or (B) a fixed amount of plate-bound antibody...
Abstract | Full Text | PDF (2363K)
Assay of Mammalian S-Adenosylmethionine Decarboxylase Activity
Author(s): Lisa M. Shantz, Anthony E. Pegg
Pub. Date: Oct-10-1997; DOI:10.1385/0-89603-448-8:45
Summary: µL 2. Set up assay tubes in a test tube rack on melting ice. Add 150 µL of the above reaction mix to each tube. 3. The reaction is started by the addition of enzyme. Add 100 µL of enzyme solution...
Abstract | Full Text | PDF (483K)
Glutaraldehyde in Protein Immobilization: A Versatile Reagent
Author(s): Lorena Betancor, Fernando López-Gallego, Noelia Alonso-Morales, Gisella Dellamora, Cesar Mateo, Roberto Fernandez-Lafuente, Jose M. Guisan
Pub. Date: Mar-15-2006; DOI:10.1007/978-1-59745-053-9_5
Summary: ). 2. Gently stir at 25°C. 3. Withdraw aliquots from suspension and supernatant and assay their catalytic activity until total adsorption of the enzyme. 4. Wash the adsorbed enzyme thoroughly...
Abstract | Full Text | PDF (150K)
Purification of DNA-Dependent RNA Polymerase from Eubacteria
Author(s): N. W. Scott
Pub. Date: Aug-30-1988; DOI:10.1385/0-89603-126-8:135
Summary: × the void volume; this can be estimated from the A 280 nm of the fractions. Use PAGE and the enzyme assay to identify the fractions containing the enzyme. 2. Pool the enzyme containing fractions...
Abstract | Full Text | PDF (841K)
Assay for G Protein-Dependent Activation of Phospholipase C β Using Purified Protein Components
Author(s): Mousumi Ghosh, Alan V. Smrcka
Pub. Date: Sept-15-2003; DOI:10.1385/1-59259-430-1:67
Summary: ; Enzyme Solution (see Note 5 ) 1. The PLCβ enzyme is used at a final concentration of 1–10 ng/reaction, depending on the type of the assay. Determine the number of reactions...
Abstract | Full Text | PDF (105K)
Lectin/Antibody “Sandwich” ELISA for Quantification of Circulating Mucin as a Diagnostic Test for Pancreatic Cancer
Author(s): Neil Parker
Pub. Date: Sept-23-1997; DOI:10.1385/0-89603-396-1:249
Summary: for the WGA coating, using diluents that had been shown to work in enzyme-linked lectin binding assays (ELLA) (10) Serum from known pancreatic cancer patients and normals were run in the assay...
Abstract | Full Text | PDF (335K)
Degradation of Chondroitin Sulfate and Dermatan Sulfate with Chondroitin Lyases
Author(s): MarÍa José Hernáiz, Robert J. Linhardt
Pub. Date: June-01-2001; DOI:10.1385/1-59259-209-0:363
Summary: . Thaw and assay activity of a frozen aliquot of enzyme (see Subheading 3.3 .). 3. Add 40 µL of Tris-HCl/sodium acetate buffer containing CS/DS sample to 10 µL of chondroitin lyase solution in a 500...
Abstract | Full Text | PDF (120K)
Enzymatic Assays to Compare Calmodulin Isoforms, Mutants, and Chimeras
Author(s): Michael P. Walsh, Jacquelyn.E. Van Lierop, Cindy Sutherland, Ritsu Kondo, J. David Johnson
Pub. Date: Jan-24-2002; DOI:10.1385/1-59259-184-1:339
Summary: . Count the cocktails in a scintillation counter (see Note 10 ). 12. Determine enzyme activity from the specific activity of the radioactive substrate. 3.7 MLCK Assay MLCK activity is absolutely...
Abstract | Full Text | PDF (255K)
Overview of Antigen Detection Through Enzymatic Activity
Author(s): Gary L. Bratthauer
Pub. Date: Jan-15-1999; DOI:10.1385/1-59259-213-9:181
Summary: oxidase. A glucose oxidase system can provide a sensitive and specific assay if other endogenous enzyme activity is a problem. 3 Chromogenic Substrates In addition to the many enzyme systems available...
Abstract | Full Text | PDF (94K)
Measurement of β-Carotene 15,15′-Dioxygenase Activity by Reverse-Phase HPLC
Author(s): Alexandrine During, Akihiko Nagao, James Cecil Smith
Pub. Date: Apr-04-2002; DOI:10.1385/1-59259-173-6:233
Summary: complicated and time-consuming steps; thus, this methodology will be useful for further investigations of β-carotene metabolism (8). This enzyme assay is described here. 24.2 Materials 24.2.1 Equipment...
Abstract | Full Text | PDF (266K)
Assay of Dolichyl-Phospho-Mannose Synthase Reconstituted in a Lipid Matrix
Author(s): John S. Schutzbach
Pub. Date: Sept-15-2006; DOI:10.1385/1-59745-167-3:31
Summary: ). Enzyme Assay in Detergent Solution 1. 0.6 mg of Dol-P (American Radiolabeled Chemicals Inc., St. Louis, MO) in CHCl3:CH3OH (2:1). 2. GDP-[3H]mannose (50 µCi; 5–15 Ci/mmol, American Radiolabeled...
Abstract | Full Text | PDF (529K)
A H-Indicator-Based Screen for Hydrolytic Haloalkane Dehalogenase
Author(s): Huimin Zhao
Pub. Date: May-16-2003; DOI:10.1385/1-59259-396-8:213
Summary: assay plate is needed. Variants with higher specific activity than the parental enzyme are selected for the next round of evolution. Fig. 1. Schematic representation of the screening method for directed...
Abstract | Full Text | PDF (151K)
Quantification of Angiotensin-Converting Enzyme (ACE) Activity
Author(s): Qing Cheng Meng, Kathleen H. Berecek
Pub. Date: Oct-29-2000; DOI:10.1385/1-59259-087-X:257
Summary: HHL and quantitates the product HA by UV detection at 228 nm. The active site-specific ACE inhibitor captopril is used to inhibit the enzyme in blank samples and increase the specificity of the assay...
Abstract | Full Text | PDF (107K)
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