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:2/4/2016)... (PRWEB) , ... February 04, ... ... conference presented by Bloomsburg University’s Digital Forensics Club, takes place February 5-6 ... two-day event features 20+ speakers and activities such as workshops and competitions ...
(Date:2/4/2016)... and MENLO PARK, Calif. , ... DMPI) ("DelMar" and the "Company"), a biopharmaceutical company focused on ... that it will present at the 18 th ... 8, 2016 at 10:00 a.m. EST in New ... DelMar,s president and CEO, will provide an update on the ...
(Date:2/4/2016)...  Spherix Incorporated (Nasdaq: SPEX ) -- an intellectual property ... intellectual property, today provided an update on the Company,s ... of Texas and announcing that ... Partes Re-examination ("IPR") proceedings that VTech and Uniden ... initiated on only certain claims of two of the ...
(Date:2/4/2016)... , February 4, 2016 ... Laboratories (ABL), Inc. --> Strasbourg, France ... --> PharmaVentures is pleased to announce that it ... its biopharmaceutical manufacturing unit in Strasbourg, France ... --> --> Transgene (Euronext: ...
Breaking Biology Technology:
(Date:2/2/2016)... --Technology Enhancements Accelerate Growth of X-ray Imaging This ... computed radiography markets in Thailand , ... (TIM). It provides an in-depth analysis of ... regional market drivers and restraints. The study offers revenue ... attractiveness, both for digital and computed radiography. Market participants ...
(Date:2/1/2016)... , Feb. 1, 2016  Wocket® smart wallet ( www.wocketwallet.com ) announces ... personality, Joey Fatone . Las Vegas , ... --> Las Vegas , where Joey appeared ... The new video ad was filmed at the Consumer Electronics Show ... the Wocket booth to meet and greet fans. ...
(Date:1/22/2016)... DUBLIN , January 22, 2016 ... has announced the addition of the  ... to their offering. --> ... of the  "Global Behavioral Biometric Market ... --> Research and Markets ( http://www.researchandmarkets.com/research/4lmf2s/global_behavioral ...
Breaking Biology News(10 mins):