Navigation Links
Preparative Nondenaturing Gel Electrophoresis Used in the Purification of an Esterase Involved in Insecticide Resistance, Rev C

on was determined empirically by analytical, nondenaturing PAGE using the Mini-PROTEAN II cell (Figure 1).

Native PAGE separates proteins both by charge and size and we had protein bands above and below our A2 protein band of interest. At each gel concentration used, the distance was measured between the band of interest and the nearest bands above and below it. The gel concentration was 9%T/3%C where the two curves intersect (Figure 1). The electrode running buffer, in both the upper and the lower buffer chambers, and the buffer used to prepare the gel, was 0.1 M Tris-borate (pH 8.0) containing 2.0 mM EDTA. The sample, after elution from hydroxyapatite, was concentrated using a Centriprep 10 concentrator (Amicon) to approximately 0.5 ml. The sample remained in the elution buffer from the hydroxyapatite column, that is, approximately 60 mM phosphate buffer (pH 6.8). The concentrated sample was mixed with a 5x solution of glycerol and marker dye Xylene Cyanole FF to give a final concentration of 10% glycerol and 0.02% dye marker. The sample, approximately 1.6 mg of protein, was loaded on a 9%T/3%C, 6 cm gel in the Model 491 prep cell (28 mm ID). The Model 491 prep cell had previously been equilibrated overnight in a 4C coldroom. Electrophoresis was performed at 15 W constant power (350400 V). Elution (1 ml/min) and fraction collection (5 ml fractions) began when the marker dye eluted from the gel, after about 1.5 hr run time (Figure 2). The enzyme activity peak was completely eluted about 3.75 hr later. The total run time was 5.5 hr. Electrode buffer was also used as elution buffer because the enzyme is stable in this buffer.


Results '"/>

Source:


Page: All 1 2 3

Related biology technology :

1. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
2. Detection of Platinum Species in Plant Material by Preparative Isotachophoresis Using the Model 491 Prep Cell, Rev A
3. Preparative Nondenaturing Gel Electrophoresis to Purify NADP-Specific Glutamate Dehydrogenase From Chlorella, Rev B
4. Preparative SDS-PAGE Electrophoresis of a Recombinant Epstein-Barr Virus Encoded Protein and Its Application in Serodiagnostic Test Systems, Rev C
5. Purification and Characterization of beta-Lactoglobulin Genetic Variants A and B Using Preparative Electrophoresis and Isoelectric Focusing
6. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
7. Semi Preparative Purification of Proteins
8. A Versatile Power Supply for All Electrophoresis Applications
9. A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis
10. Continuous-Elution Electrophoresis for Purification of the Baculovirus-Expressed Coronavirus Structural Proteins, Rev A
11. Detection of K-ras Point Mutations in the Pancreas by Constant Denaturing Gel Electrophoresis Using the DCode System
Post Your Comments:
(Date:8/26/2015)... ... August 26, 2015 , ... PRC Clinical, a Clinical ... relationships with local sponsors in southern California. The Clinical Trial Management Expert CRO seeks ... and regulatory pathways., , Stem Cell Meeting on the ...
(Date:8/25/2015)... 25, 2015   WuXi PharmaTech (Cayman) Inc. (NYSE: ... technology platform company serving the pharmaceutical, biotechnology, and medical ... Hong Kong Exchange Stock Code: 0950), a research-based biopharmaceutical ... with 20 years of operations in China ... signed an agreement whereby WuXi,s Laboratory Testing Division (LTD) ...
(Date:8/25/2015)... Menomonie, Wis. (PRWEB) , ... August 25, 2015 ... ... and fermentor at its research facility in Knapp, Wis. The company has added ... Biostat C15 Fermentor allows detailed data collection during bacterial or fungal fermentation process ...
(Date:8/25/2015)... ... 2015 , ... One of the world’s foremost authorities on supply chain risk ... been on a mission for the past two weeks. She’s been scrambling to warn ... the two massive explosions that occurred on August 12 at the Ruihai International Logistics ...
Breaking Biology Technology:PRC Clinical To Attend Major Industry Conferences And Meet Sponsor Companies In The San Diego Biotech Hub 2PRC Clinical To Attend Major Industry Conferences And Meet Sponsor Companies In The San Diego Biotech Hub 3WuXi PharmaTech to Be Exclusive Supplier of Laboratory Testing Services to Lee's Pharm 2WuXi PharmaTech to Be Exclusive Supplier of Laboratory Testing Services to Lee's Pharm 3New centrifuge and fermentor expand product development capacity at Vets Plus 2Resilinc Supply Chain Expert Warns: Impact of Tianjin Chemical Explosions Will be Far Worse than Reported 2Resilinc Supply Chain Expert Warns: Impact of Tianjin Chemical Explosions Will be Far Worse than Reported 3Resilinc Supply Chain Expert Warns: Impact of Tianjin Chemical Explosions Will be Far Worse than Reported 4
... the Board of Directors,meeting held today, the Directors ... The dividend is payable on December 15, 2008 ... About Sigma-Aldrich: Sigma-Aldrich is a leading Life Science ... products and kits,are used in scientific and genomic ...
... 11 /Xinhua-PRNewswire-FirstCall/ -- BMP,Sunstone Corporation (Nasdaq: BJGP ... due to the technical difficulties encountered on,today,s conference ... tomorrow,Wednesday, November 12, 2008. On this call, BMP ... since technical difficulties prevented the,Company from addressing all ...
... molecule detection to expand prominent Scientific Advisory ... ... 11 Quanterix Corporation, the,leader in single molecule detection, announced today ... Corn is a,Professor in the Departments of Chemistry and Biomedical Engineering ...
Cached Biology Technology:BMP Sunstone to Host Follow-Up 3Q08 Conference Call Due to Technical Difficulties Encountered on Tuesday's Call 2BMP Sunstone to Host Follow-Up 3Q08 Conference Call Due to Technical Difficulties Encountered on Tuesday's Call 3Quanterix Corporation Announces the Addition of Dr. Robert Corn to its Scientific Advisory Board 2
(Date:8/6/2015)... 2015 Synaptics Inc. (NASDAQ: SYNA ... today announced collaboration with Microsoft on biometric fingerprint ... leveraged Synaptics, deep expertise in human interface which ... Through stringent testing, Synaptics, advanced image sensing ... Precision TouchPad (PTP) specification empowering OEMs with Windows ...
(Date:8/4/2015)... (NASDAQ: AMRI ) today reported financial and operating results for ... , Second quarter contract revenue of $ 85.2 ... Adjusted contract margins of 26 % , ... in EPS from royalties in the current quarter , ... are very pleased to present another strong financial quarter, with all ...
(Date:7/31/2015)... La 10e Conférence internationale sur la ... le BGI du 22 au 25 octobre 2015 ... La conférence célèbre son 10e anniversaire ... est devenue l,une des réunions annuelles mondiales les ... c,est aussi la plus dynamique, la plus enthousiaste ...
Breaking Biology News(10 mins):Synaptics Chosen for Human Interface Co-Development on Windows 10 2AMRI Announces Second Quarter 2015 Results 2AMRI Announces Second Quarter 2015 Results 3AMRI Announces Second Quarter 2015 Results 4AMRI Announces Second Quarter 2015 Results 5AMRI Announces Second Quarter 2015 Results 6AMRI Announces Second Quarter 2015 Results 7AMRI Announces Second Quarter 2015 Results 8AMRI Announces Second Quarter 2015 Results 9AMRI Announces Second Quarter 2015 Results 10AMRI Announces Second Quarter 2015 Results 11AMRI Announces Second Quarter 2015 Results 12AMRI Announces Second Quarter 2015 Results 13AMRI Announces Second Quarter 2015 Results 14AMRI Announces Second Quarter 2015 Results 15AMRI Announces Second Quarter 2015 Results 16AMRI Announces Second Quarter 2015 Results 17AMRI Announces Second Quarter 2015 Results 18AMRI Announces Second Quarter 2015 Results 19AMRI Announces Second Quarter 2015 Results 20AMRI Announces Second Quarter 2015 Results 21AMRI Announces Second Quarter 2015 Results 22AMRI Announces Second Quarter 2015 Results 23AMRI Announces Second Quarter 2015 Results 24La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 2La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 3
... Boston University School of Medicine (BUSM) have identified a ... response and homeostasis. These findings could help lead to ... associated with cancer, type 2 diabetes and other diseases. ... PhD, assistant professor of biochemistry at BUSM, was done ...
... This press release is available in German . ... is a key feature of human evolution. In particular, humans ... individuals. Chimpanzees and orangutans, two of our closest living relatives, ... generation to another. Whether and how this process resembles the ...
... a warning for obese women who are planning pregnancies. Even if ... will develop in an unhealthy environment that places the infants at ... in the placentas of obese mothers when it should be going ... in the placenta of an obese mother is half the size ...
Cached Biology News:BUSM researchers identify key regulator of inflammatory response 2Majority-biased learning 2U of I study: Lose body weight before gaining baby weight 2
Request Info...
... StabilZyme HRP Conjugate Stabilizer is an ... protein and other non-toxic stabilizing chemicals in ... 6.2 6.7. This product contains ... and 20 ppm Proclin 300 (Rohm and ...
Preparation Note: by a modification of the procedure of C.W. Hancher et al., Biotechnol. Bioeng. 11, 1055 (1969) Assay: ~ 20 A 260 /mg Solubility: water 10 mg/mL, clear, light-yellow Suitab tst...
Request Info...
Biology Products: