Navigation Links
A Microtiter-based Assay for the Determination of ID50s of b-lactamase Inhibitors Employing Reporter Substrates Detected at UV or Visible Wavelengths (MaxLine Application Note #20)

es the UV capability of the SPECTRAmax 250 micro-plate spectrophotometer and can be readily modified to determine ID50 values of inhibitors of other enzymes requiring UV/Vis monitoring.


MATERIALS AND METHODS

Materials
1. SPECTRAmax 250 microplate spectrophotometer
2.SOFTmax PRO software
3.Quartz microplates
4. Deep well microplate
5. Multichannel pipette
6. DMSO
7. Imipenem and nitrocefin
8. β -lactamase preparation
9. Experimental compounds

Method for determining ID50 values following a 5 minute incubation of enzyme and inhibitor.

Step 1 Set the incubator temperature to 37C., then allow it to equilibrate for at least 30 minutes

Step 2 Set up SOFTmax PROs Instrument Settings dialog box for imipenem as shown in Figure 1 and for nitrocefin as shown in Figure 2.



Step 3 Dissolve the inhibitors in 25 mM piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES) buffer, pH 7.0 with no added Zn2+ to produce a 3 mM solution. 5% DMSO can be used if needed to aid in dissolving the inhibitors.

Step 4 In a deep-well 96 well plate, pipette 600 l of 25 mM PIPES buffer, pH 7.0 into each well in columns 1-11.

Step 5 Pipette 1 mL 3 mM inhibitor solution into each well in column 12.

Step 6 Using a multichannel pipette, make a 1:3 serial dilution of the 3 mM solution down to column 3: take 300 l of 3 mM solution from column 12 and dispense into column 11. Mix, then take 300 l of column 11 and dispense into column 10, and so on across the columns to column 3 which will then be 0.15 M.

Step 7 Aliquot 50 l of the contents of the deep wells into the corresponding wel
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. A Positive Selection Assay for Mutation Analysis in Big Blue Animals
2. Luminol-enhanced Assay for Superoxide Anion (O2 face="Symbol" )
3. Organize Your Data from the Big Blue Mutation Assay
4. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
5. RNase Protection Assay (RPA) Using DIG-Labeled RNA Probes
6. measuring cell proliferation with SpectraMax M5, M2 and Gemini microplate readers using the CyQUANT Cell Proliferation Assay
7. Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success
8. Using Validated siRNAs in Functional Genomic Assays
9. Nuclease Protection Assays: The Basics
10. RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay
11. Detection of Phosphorylated and Total ERK and p38 MAPK by Bio-Plex Assay, ELISA, and Western Blotting, Rev A
Post Your Comments:
(Date:9/15/2014)... ANNAPOLIS, Maryland , 15. September 2014 /PRNewswire/ ... Week (IPAW) wird vom 12. bis 18. ... der Plasma Protein Therapeutics Association (PPTA) ... Ziele: , Das globale Bewusstsein für ... Den Beitrag von Plasmaspendern beim Retten und Verbessern ...
(Date:9/15/2014)... Sept. 15, 2014  The second annual International ... 12-18. A joint initiative of the Plasma Protein ... , IPAW is designed to: , Raise ... the contributions of plasma donors in saving and improving ... therapies and rare diseases Member companies ...
(Date:9/14/2014)... protein functions in a wide variety of ways, including ... protein can take on another form and perform no ... fold wrongly can cause serious diseases such as Alzheimer,s, ... have a tendency to "infect" other molecules of the ... or plaques. These amyloids can damage cells and tissues ...
(Date:9/12/2014)... a magnet using an electric field (magnetoelectric memory ... the next-generation technology for information processing and storage. ... effect, due to the coexistence of electric and ... of spontaneous electric and magnetic polarizations is rare ... of the MEM effect. This article briefly reviews ...
Breaking Biology Technology:International Plasma Awareness Week feiert Spender und rückt seltene Krankheiten in den Mittelpunkt 2International Plasma Awareness Week feiert Spender und rückt seltene Krankheiten in den Mittelpunkt 3International Plasma Awareness Week To Celebrate Donors And Spotlight Rare Diseases 2International Plasma Awareness Week To Celebrate Donors And Spotlight Rare Diseases 3Measuring modified protein structures 2Measuring modified protein structures 3
... SOUTH SAN FRANCISCO, Calif., June 11 iZumi ... industrialization of induced pluripotent stem cell (iPS) technology ... cellular reprogramming and directed differentiation of patient cells, ... Ph.D., to the company,s board of directors and ...
... Calif., June 11 STAAR Surgical Company (Nasdaq: ... of minimally invasive ophthalmic products, today announced that the ... System for use with the Affinity(TM) Collamer(R) Three-Piece NTIOL ... Epiphany will usher in a new era of insertion ...
... WALTHAM, Mass., June 11 Repligen Corporation (Nasdaq: ... fiscal year 2009, ended March 31, 2009. Total revenue ... of $19,296,000 for fiscal year 2008 ended March 31, 2008, ... the year was comprised of Protein A product revenue, and ...
Cached Biology Technology:iZumi Bio Appoints Dr. Corey Goodman to Board of Directors and Scientific Advisory Board 2iZumi Bio Appoints Dr. Corey Goodman to Board of Directors and Scientific Advisory Board 3STAAR Surgical Receives FDA Clearance for the Epiphany(TM) Injector System 2STAAR Surgical Receives FDA Clearance for the Epiphany(TM) Injector System 3Repligen Reports Fourth Quarter and Fiscal Year 2009 Financial Results 2Repligen Reports Fourth Quarter and Fiscal Year 2009 Financial Results 3Repligen Reports Fourth Quarter and Fiscal Year 2009 Financial Results 4Repligen Reports Fourth Quarter and Fiscal Year 2009 Financial Results 5Repligen Reports Fourth Quarter and Fiscal Year 2009 Financial Results 6Repligen Reports Fourth Quarter and Fiscal Year 2009 Financial Results 7
(Date:9/16/2014)... make ethical choices such as buying clothing not made ... coffee, and bringing their own bags when they go ... Journal of Consumer Research , ethical consumption is motivated ... about unethical practices into action. , "Advocates of ethical ... human costs of the products they choose, but unfortunately ...
(Date:9/16/2014)... N.C. , Sept. 16, 2014  Valencell, Inc., ... been chosen as one of 18 "Showcase Companies" representing ... businesses at the annual CED Tech Venture Conference on ... Convention Center in Raleigh, North ... will lead a discussion during the "Digital Health ...
(Date:9/16/2014)... a season of bounty. Fungi of dizzying variety erupt ... in search of elusive but delectable wild mushrooms. Part ... treasure hunt, and their mysterious not-quite-meat, not-quite-vegetable qualities that ... which you are eating known to science? , The ... although some estimates range up to 10 million species, ...
Breaking Biology News(10 mins):Why are consumers willing to spend more money on ethical products? 2Valencell, Inc. Selected as 'Showcase Company' at CED Tech Venture Conference 2014 2What's for dinner? Rapidly identifying undescribed species in a commercial fungi packet 2What's for dinner? Rapidly identifying undescribed species in a commercial fungi packet 3
... disease are increasingly becoming resistant to synthetic insecticides. Now ... Complementary and Alternative Medicine suggests that the berries of ... have potential for keeping mosquitoes at bay. S. ... for its medicinal properties and commonly used as ...
... Southern Research Station (SRS), Iowa State University, and the ... of a fungus responsible for the wilt of redbay ... Florida. In the February issue of Plant Disease, ... results from their assessment of the fungus, the beetle ...
... but the microscopic animals called bdelloid rotifers seem to get ... have done so over millions of years of evolution, resulting ... the usual drawback of asexuality extinction and the ... finding out how. In two related papers published ...
Cached Biology News:Ayurvedic nightshade deadly for dengue mosquito 2Laurel wilt of redbay and sassafras: Will avocados be next? 2Is DNA repair a substitute for sex? 2Is DNA repair a substitute for sex? 3
...
Request Info...
Request Info...
... KingFisher magnetic particle based purification systems provide ... from any sample. They offer increased throughput ... three systems- KingFisher, KingFisher mL and KingFisher ... 1000 l. Each system consists of an ...
Biology Products: