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.

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


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:

(Date:11/25/2015)... and HOLLISTON, Mass. , ... Inc. (Nasdaq: HART ), a biotechnology company developing ... CEO Jim McGorry will present at the ... 1, 2015 at 2:30 p.m. PT. The presentation will ... for 30 days. Management will also be available at ...
(Date:11/25/2015)... The Global Genomics Industry ... and in-depth study on the current state of ... ) , The report provides ... classifications, applications and industry chain structure. The Genomics ... including development trends, competitive landscape analysis, and key ...
(Date:11/24/2015)... 2015 Halozyme Therapeutics, Inc. (NASDAQ: HALO ) will ... New York on Wednesday, December 2 at 9:30 a.m. ... and CEO, will provide a corporate overview. th ... at 1:00 p.m. ET/10:00 a.m. PT . Jim Mazzola ... a corporate overview. --> th Annual Oppenheimer Healthcare ...
(Date:11/24/2015)... RALEIGH, N.C. , Nov. 24, 2015  Clintrax Global, Inc., ... Raleigh, North Carolina , today announced that the company has ... earnings represented a 391% quarter on quarter growth posted for Q3 ... Kingdom and Mexico , with the ... place in December 2015. --> United Kingdom ...
Breaking Biology Technology:
(Date:10/23/2015)... 23, 2015 Research and Markets ( ... Voice Recognition Biometrics Market 2015-2019" report to their ... The global voice recognition biometrics market to grow ... --> --> The report, ... based on an in-depth market analysis with inputs from ...
(Date:10/22/2015)... AWRE ), a leading supplier of biometrics software and services, ... 2015.  --> --> Revenue ... of 33% compared to $6.0 million in the same quarter last ... million, or $0.10 per diluted share, which compared to $2.6 million, ... ago.  --> --> Lower ...
(Date:10/22/2015)... NEW YORK , Oct. 22, 2015 ... matches the fingerprint scan of an individual with the ... features such as arch, whorl, and loop. Pattern-based algorithms ... database. With advances in technology, AFIS was introduced in ... law enforcement agencies to identify a criminal. ...
Breaking Biology News(10 mins):