Single-step electrotransfer of reverse-stained proteins from sodium dodecyl sulfate-polyacrylamide gel onto reversed-phase minicartridge and subsequent desalting and elution with a conventional high-performance liquid chromatography gradient system for analysis

1995 ◽  
Vol 16 (1) ◽  
pp. 911-920 ◽  
Author(s):  
Carlos Fernandez-Patron ◽  
Joel Madrazo ◽  
Eugenio Hardy ◽  
Enrique Mendez ◽  
Rainer Frank ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Sodium Dodecyl Sulfate ◽  
High Performance ◽  
Sodium Dodecyl ◽  
Reversed Phase ◽  
Single Step ◽  
Polyacrylamide Gel ◽  
Gradient System ◽  
Dodecyl Sulfate

SELECTION OF COLUMN AND OPERATING CONDITIONS FOR REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PROTEINS IN CANADIAN WHEAT

1985 ◽  
Vol 65 (2) ◽  
pp. 285-298 ◽  
Author(s):  
J. E. KRUGER ◽  
B. A. MARCHYLO
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Storage Proteins ◽  
Sodium Dodecyl ◽  
Reversed Phase ◽  
Operating Conditions ◽  
Protein Patterns ◽  
Optimal Separation ◽  
Rp Hplc

Chromatographic conditions were optimized and three commercially available columns were evaluated for separation of alcohol-soluble storage proteins of Neepawa wheat using reversed-phase high-performance liquid chromatography (RP-HPLC). Optimal separation was achieved using an extracting solution of 50% 1-propanol, 1% acetic acid, and 4% dithiothreitol and an HPLC elution time of 105 min at a flow rate of 1.0 mL/min. HPLC columns evaluated (SynChropak RP-P, Ultrapore RPSC and Aquapore RP-300) varied in selectivity and resolution. The column providing the greatest versatility was Aquapore RP-300 available in cartridge form. Sodium dodecyl sulfate gradient-gel electrophoresis analysis of protein peaks resolved by RP-HPLC indicated that many of the eluted peaks contained more than one protein species. Chromatographic protein patterns obtained for Neepawa wheat grown at different locations and in different years were qualitatively the same.Key words: Protein, high-performance liquid chromatography, wheat

scholarly journals Removal of sodium dodecyl sulfate from protein samples

Chemija ◽  
2018 ◽  
Vol 29 (4) ◽  
Author(s):  
Andrius Žilionis
Keyword(s):  
Liquid Chromatography ◽  
Sodium Dodecyl Sulfate ◽  
Protein Denaturation ◽  
Sodium Dodecyl ◽  
Reversed Phase ◽  
Spectrometric Analysis ◽  
Photometric Method ◽  
Ionization Mass ◽  
Dodecyl Sulfate ◽  
Model Protein

Sodium dodecyl sulfate (SDS) is a widely used detergent for protein denaturation and solubilization. However, application of SDS in the sample preparation for the liquid chromatographic-mass spectrometric analysis is limited because commonly used SDS concentrations interfere with reversed phase liquid chromatography and electrospray ionization mass spectrometry. In order to analyse SDS pretreated proteins by the above-mentioned methods SDS must be completely removed or its concentration must be lowered to less than 0.01%. In this work we present a comparison of different SDS removal strategies based on SDS ultrafiltration, protein precipitation and SDS precipitation methods. Every strategy was optimized so that the initial 4% SDS concentration was lowered to less than 0.01% and the initial sample volume remained unchanged. The modified Mukerjee’s photometric method was used for the SDS quantitation in the presence of model protein bovine serum albumin and the recovery of model protein was evaluated using reversed phase ultra performance liquid chromatography. The main advantages and drawbacks of every strategy are discussed.

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