Navigation Links
Semi Preparative Purification of Proteins


Most Basic Recommended Cecil Instrumentation:
Cecil Adept HPLC System 4BP, semi-preparative, biocompatible with a PEEK 500 l loop
Fraction collector
Column heater


Introduction:
Researchers need to understand the expression of proteins. They also would like to understand protein expression in healthy and diseased cells.

Hence the study of proteomics, which characterises and analyses of the function of proteins in the body is crucial.

In general, a mixture of proteins is separated by HPLC the resultant major peaks are collected in a fraction collector. This collected material is then suitable for characterisation by amino acid analysis, sequence analysis, capillary electrophoresis or SDS-PAGE gel electrophoresis.

The same analytical HPLC method may be used to elute a given protein mixture. If larger quantities of the separated proteins are required, the user can very easily scale up the analytical method to semi-preparative scale. The key parameter for transferring an HPLC separation from the analytical to preparative scale is column volume. Both mass load and volume load need to be scaled in proportion to column volume. Flow rates should be scaled up in proportion to column volumes.

The HPLC method may for example, involve the use of ion exchange , gel filtration or affinity chromatography techniques, depending on the proteins of interest A uv/visible HPLC detector is often used.

If there is a limited quantity of sample, reverse phase narrow bore HPLC column (0.21 x 25cm) is useful. Analytical HPLC reverse phase columns, 0.46 x 25cm, are useful for larger sample quantities up to 500g. Semi-preparative HPLC , to 3mg/run, may be run on wider bore columns such as 2.2 x 25cm.

In some cases, it may be necessary to remove interfering and abundant proteins, such as albumin, IgG, IgA, transferrin, haptoglobin, and antitrypsin and dye terminators from human serum sample matrices. This may be performed by the use of Multiple Affinity Removal Systems or by the use of Spin columns. These systems and columns are used to pre-treat samples, before they are injected onto an HPLC column.

Samples should not be suspended in detergent or non volatile buffer containing solutions. Some detergents may bind to reverse phase columns and modify them irreversibly. Samples should not be dissolved in organic solvents, as this may inhibit the proteins from adsorbing to the stationary phase.


Examples:





back to top
'"/>

Source:


Page: All 1 2

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 Used in the Purification of an Esterase Involved in Insecticide Resistance, Rev C
4. Preparative Nondenaturing Gel Electrophoresis to Purify NADP-Specific Glutamate Dehydrogenase From Chlorella, Rev B
5. Preparative SDS-PAGE Electrophoresis of a Recombinant Epstein-Barr Virus Encoded Protein and Its Application in Serodiagnostic Test Systems, Rev C
6. Purification and Characterization of beta-Lactoglobulin Genetic Variants A and B Using Preparative Electrophoresis and Isoelectric Focusing
7. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
8. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
9. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
10. Yeast Protein Production System Features High Yields and One-Step Purification
11. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... ... 23, 2016 , ... Charm Sciences, Inc. is pleased to ... AOAC Research Institute approval 061601. , “This is another AOAC-RI approval of the ... Vice President of Regulatory and Industrial Affairs. “The Peel Plate methods perform comparably ...
(Date:6/23/2016)... ... June 23, 2016 , ... Supplyframe, ... of the Supplyframe Design Lab . Located in Pasadena, Calif., the Design ... of how hardware projects are designed, built and brought to market. , The ...
(Date:6/23/2016)... BEACH, Calif. , June 23, 2016  Blueprint ... new biological discoveries to the medical community, has closed ... co-founder Matthew Nunez . "We have ... us with the capital we need to meet our ... will essentially provide us the runway to complete validation ...
(Date:6/23/2016)... Prairie, WI (PRWEB) , ... June 23, 2016 ... ... consultancy focused on quality, regulatory and technical consulting, provides a free webinar ... is presented on July 13, 2016 at 12pm CT at no charge. , ...
Breaking Biology Technology:
(Date:6/9/2016)... , June 9, 2016 ... Police deploy Teleste,s video security solution to ensure the safety ... France during the major tournament Teleste, ... communications systems and services, announced today that its video security ... to back up public safety across the country. ...
(Date:6/2/2016)... NEW YORK , June 2, 2016   The ... (Weather), is announcing Watson Ads, an industry-first capability in which ... advertising, by being able to ask questions via voice or ... Marketers have long ... with the consumer, that can be personal, relevant and valuable; ...
(Date:5/20/2016)...  VoiceIt is excited to announce its new ... By working together, VoiceIt and VoicePass will offer ... take slightly different approaches to voice biometrics, collaboration ... usability. Both ... "This marketing and technology partnership allows ...
Breaking Biology News(10 mins):