Navigation Links
New gravity-flow column gives faster and simpler purification of histidine-tagged proteins

roximately four times faster on His GraviTrap (Fig 2B).

Complete details of these comparisons are provided in data file 11-0036-90, available at www.amershambiosciences.com/promo_dm205

His GraviTrap also enables rapid separation of large proteins. To demonstrate this, His GraviTrap was used to purify (His)10-TRX-P450 (Mr 133 200). E. coli JM109 cells expressing (His)10-TRX-P450 were enzymatically and mechanically lysed and a 20 ml clarified sample was loaded onto His GraviTrap using LabMate. The whole purification took just 25 min.

Results show that the purification yielded three major protein bands when analyzed by SDS-PAGE (Fig 3A, lane 4). Western blot analysis confirms that each of the three bands contains a histidine-tag (Fig 3B). However, only a weak band can be seen from the band at approximately Mr 15 000. Some of the smaller proteins may have passed through the membrane due to a prolonged blotting time. N-terminal sequencing (data not shown) of the three bands confirms that the low molecular weight bands are truncated forms of the histidine-tagged target protein. To separate the full-length protein from the truncated forms, a second purification step, such as gel filtration, is recommended.


Summary
His GraviTrap columns provide a very fast and simple method for purifying histidine-tagged proteins without the need for a pump or purification system. The prepacked Ni Sepharose 6 Fast Flow gives high protein binding capacity. Proven protocols, application data, and new support products help optimize purification results and simplify the whole procedure.

To obtain data file 11-0036-90, visit www.am
'"/>


Source:


Page: All 1 2 3 4

Related biology technology :

1. The combination of monolithic columns and modern HPLC/MS/MS instrumentation for high chromatographic throughput and sensitive detection of beta-agonists
2. TSKgel ODS-100Z and TSKgel ODS-100V column series provides a universal solution for the analysis of samples containing acidic, basic, polar & non-polar compounds
3. Fully automated DNA purification and efficient multiplex PCR for analysis of microsatellite loci
4. Laser microdissection and nucleic acid purification - a Leica - QIAGEN cooperation
5. DirectPrep 96 Miniprep System for cost-effective, high-throughput plasmid DNA purification
6. Ni-NTA Superflow Columns - automated large-scale purification of 6xHis-tagged proteins
7. Walkaway automation for nucleic acid purification
8. DNA purification from pig ears to bird blood using proven DNeasy Technology
9. Addition of imidazole during binding improves purity of histidine-tagged proteins
10. 2100 expert software Powerful software for the analysis of RNA, DNA, proteins, and cells with the Agilent 2100 bioanalyzer
11. Application of the Bio-Plex Cell Lysis Kit in Western Blot Analysis of Phosphoproteins, Rev A
Post Your Comments:
(Date:10/31/2014)... UK (PRWEB) October 30, 2014 Caustic soda ... epoxy resins and polycarbonates. At present, China takes lead in ... , Dow, which possesses a number of facilities in the ... in the world caustic soda market in terms of output ... forecast to grow on average by 3% per year in ...
(Date:10/31/2014)... Chicago, IL (PRWEB) October 30, 2014 ... tapped Insight Product Development’s decades-long heritage in successful ... real world context for what it takes to ... medical space. As part of a two-hour on-site ... University’s Kellogg school of business, McCormick School of ...
(Date:10/31/2014)... 30, 2014 Two separate delegations ... hosted by Charm Sciences at its Andover, Massachusetts ... in the US, the participants discussed international systems ... management, and process verification, and received hands-on technical ... was the first visit to the United States ...
(Date:10/31/2014)... Jeffrey T. Manuel, assistant professor in Southern Illinois ... of Arts and Sciences, is teaming up with the ... fuels in Illinois. , “Farmers, researchers, business ... build Illinois’ biofuels industry for decades,” Manuel said. “This ... agricultural and business history. , “Fuel alcohol has ...
Breaking Biology Technology:China to Dominate Caustic Soda Market Through 2018, Says Merchant Research & Consulting in Its New Study 2Insight Product Development Gives Northwestern University Students Foundation for Innovation Success 2Insight Product Development Gives Northwestern University Students Foundation for Innovation Success 3Charm Sciences Hosts Executives from Latin America for Dairy Safety Executive Seminar 2SIUE History Prof Manuel Working on Biofuels Book with NCERC 2SIUE History Prof Manuel Working on Biofuels Book with NCERC 3
... Prepare DNA from blood, cultured cells, or bacteria: , ... boil for 8 min. , ... substrate for PCR amplification. , DNA Preparation From ... frozen, refrigerated, or collected fresh. , 1. Add 36 l ...
... , AquaPure Genomic DNA Isolation Kit , ... , Method , ... tissue sample quickly and freeze in liquid nitrogen. Store , at ... keep tissue on ice at all times including when tissue is weighed. , ...
... AquaPure Genomic DNA Isolation Kit , Catalog #732-6340 , ... Method , Cell Lysis ... to avoid clotting. Alternatively, , collect whole blood in EDTA to ... may be either fresh or frozen. Store fresh blood , sample ...
Cached Biology Technology:InstaGene Matrix: Prepare DNA Templates for PCR With No Phenol/Chloroform Extractions and No Deproteinization Steps, Rev C 2Genomic DNA Isolation From 510 mg Marine Invertebrate Tissue 2Genomic DNA Isolation Protocol for 40 l Avian Whole Blood 2
(Date:10/31/2014)... Melbourne researchers have uncovered key steps involved in programmed ... diseases including lupus, cancers and neurodegenerative diseases. , ... Institute worked together to discover the three-dimensional structure of ... the first steps in how it causes cell death. ... and Proceedings of the National ...
(Date:10/31/2014)... A University of Adelaide-led project has developed a new ... vaccinated against the H5N1 strain of avian influenza virus ... infected. , This is a significant step in the ... which is widespread in the poultry populations of South ... global concern because of its possible transmission to humans ...
(Date:10/31/2014)... been studying the behavior of approximately 300 wild ... They observed that ravens slowly build alliances through ... they also observed that these affiliative interactions were ... about 50 % of the cases these interventions ... ravens, intervening can be potentially risky when the ...
Breaking Biology News(10 mins):A matter of life and death: Cell death proteins key to fighting disease 2New step towards eradication of H5N1 bird flu 2
... Madison, WI, May 1, 2009 -- With each passing year, ... growth has created a wide ranging strain on our water ... unprecedented urgency to address the issue of food security. One ... the genetic modification of seeds, both as a method of ...
... latest American Chemical Society (ACS) Weekly PressPac from ... from ACS, 34 peer-reviewed journals and Chemical ... journal or the American Chemical Society as the ... ARTICLE #1 FOR IMMEDIATE RELEASE ...
... 4, 2009 Women who have babies naturally in their ... Now, a new study shows their brothers also live longer, ... prolong lifespan and female fertility, and may be more important ... family give birth at older ages, you may well have ...
Cached Biology News:Food security: It starts with seed 2American Chemical Society's weekly PressPac -- April 29, 2009 2American Chemical Society's weekly PressPac -- April 29, 2009 3American Chemical Society's weekly PressPac -- April 29, 2009 4American Chemical Society's weekly PressPac -- April 29, 2009 5American Chemical Society's weekly PressPac -- April 29, 2009 6Late motherhood boosts family lifespan 2Late motherhood boosts family lifespan 3Late motherhood boosts family lifespan 4Late motherhood boosts family lifespan 5
... reactivity with rat (100%), mouse ... pig (48%). Slight reactivity ... confirmed by Western blot versus ... Gia-1, Gia-2, rat Gia-3, bovine ...
KV beta.1 (Q-13)...
Preadipocyte factor-1 [Pref-1, DLK-1], C-terminal...
... 4 deuterium atoms at the 3, 3', ... for use as an internal standard for ... GC- or LC-mass spectrometry. PGE2 is one ... acid and one of the most widely ...
Biology Products: