Navigation Links
Purification of Insulin on UNO S1 Column, Rev A


Insulin is a peptide hormone which stimulates anabolic reactions for carbohydrates, proteins, and fats resulting in lowered blood glucose. It is used therapeutically for treatment of diabetes mellitus and was the first recombinant protein on the market. Although insulin is well characterized and understood, it is often difficult to purify because of the harsh pH and denaturing conditions needed to solubilize and disaggregate the protein. A comparison of the UNO S1 (Bio-Rad catalog # 720-0021) and a popular monodispersed cation exchange column showed that it was advantageous to use the UNO S1 column for small scale insulin separation or analysis.


Experimental
The insulin used in this study was an acid precipitate in suspension. The insulin was solubilized in acidic buffer with urea. The insulin sample was applied separately to an UNO S1 and a monodispersed cation exchange column and eluted by linear gradients. Fractions were collected and neutralized with ammonium hydroxide to pH 7.0. Due to the viscosity of the buffers, both columns were run at half the maximum recommended flow rates. The separation performed on the UNO S1 was run at 3 ml/min (Figure 1). The separation performed on the monodispersed cation exchanger was performed at 1 ml/min (Figure 2). Fractions 4350 and fractions 3034 were pooled from the separation performed on the UNO S1 and the monodispersed column respectively and analyzed by reversed phase HPLC. Analysis performed on Hi-Pore RP-304 reversed phase column (Bio-Rad catalog # 125-0551). The products from both columns were 98% pure. Figure 3 shows the HPLC chromatogram of the pooled fractions from the UNO S1 column after subtraction of the background UV from the buffer.


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
2. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
3. Yeast Protein Production System Features High Yields and One-Step Purification
4. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
5. Easy Cosmid DNA Purification with StrataPrep Kits
6. MinElute PCR Purification Kit
7. QIAquick PCR Purification Kit
8. Microarray Target Purification Kit and the Microarray Target Synthesis Kit (T7)
9. Double-Affinity Purification of GST-His Tagged Proteins Using Vivapure 8-to-96 Well Cobalt-Chelate Kit
10. Ultra-Fast Purification of Small Nucleic Acids
11. RNAClean - RNA & cDNA in vitro Reaction Purification
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:12/2/2016)... Boston, MA (PRWEB) , ... December 02, 2016 , ... ... of Light Event on December 3rd, 2016. The event, which is held on ... NTI’s work with helping Americans with Disabilities back into the workplace. Suitable Technologies is ...
(Date:12/2/2016)... The immunohistochemistry (IHC) market is projected to reach ... during the forecast period of 2016 to 2021 dominated by immunohistochemistry ... the largest share of immunohistochemistry (IHC) market, by end user.   ... , , ... across 225 pages, profiling 10 companies and supported with 181 tables ...
(Date:11/30/2016)... , ... November 30, 2016 , ... ... moving magnet Voice Coil Actuator with a flexure design that ensures high alignment ... with cost-effective pricing and is ideally suited where extreme precision is required, such ...
(Date:11/30/2016)... New York , November 30, 2016 ... consolidated as a few players hold a dominant share ... Group, Charles River Laboratories International, Inc., and Merck KGaA, ... market in 2015. Transparency Market Research observes that these ... are focused on development products that are do not ...
Breaking Biology Technology:
(Date:11/16/2016)... Sensory Inc ., a Silicon ... consumer electronics, and VeriTran , a technology ... today announced a global partnership that will provide ... users of mobile banking and mobile payments solutions.  ... which requires no specialized biometric scanners, yet provides ...
(Date:6/27/2016)... Research and Markets has announced the addition of the "Biometrics ... The report forecasts the biometrics market ... CAGR of 12.28% during the period 2016-2020. The ... inputs from industry experts. The report covers the market landscape and ... a discussion of the key vendors operating in this market. ...
(Date:6/20/2016)... Securus Technologies, a leading provider of ... safety, investigation, corrections and monitoring announced that after ... secured the final acceptance by all three (3) ... Systems (MAS) installed. Furthermore, Securus will have contracts ... by October, 2016. MAS distinguishes between legitimate wireless ...
Breaking Biology News(10 mins):