Sample preparation is often necessary to separate and concentrate various compounds prior to analysis of complex samples. In this regard, isoelectric focusing (IEF) is one of the best sample preparation methods. With this approach, however, carrier ampholytes have to be introduced into the samples, which may result in matrix interferences. In this paper, a simple ampholyte-free IEF free-flow electrophoresis design was developed for the separation of proteins. beta-Lactoglobulin, hemoglobin, myoglobin and cytochrome c were selected as model analytes. The experimental design took advantage of the electrolysis-driven production of H(+) and OH(-) ions that migrated from the anode and cathode, respectively, establishing a pH gradient spanning from 2.3 to 8.9. The separation chamber was filled with silanized glass beads as a support medium. Dialysis membranes were mounted at the two sides of the separation chamber (made of glass slides) and sealed with 2% agarose gel. The separated proteins drained from the outlets of the separation chamber and could be successfully collected into small glass tubes. The focusing process was visually observed and the separation was confirmed by capillary isoelectric focusing (cIEF) with pI markers.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.chroma.2008.07.093 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
The clinical significance of mirror patterns of cerebrospinal fluid oligoclonal immunoglobulin G bands (IgG-OCBs) in peripheral neuropathy disorders.
Clin Chim Acta
January 2025
Department of Neurology, Huashan Hospital, Fudan University and Institute of Neurology, Fudan University, National Center for Neurological Disorders, Shanghai, China. Electronic address:
Background: CSF (cerebrospinal fluid) oligoclonal immunoglobulin G bands (IgG-OCBs) analysis plays a crucial role in diagnosis of various neurological disorders. However, the clinical significance of mirror pattern bands remains unclear, and their precise application is not well understood.
Methods: We retrospectively reviewed a total of 7597 IgG-OCB records detected using isoelectric focusing from May 2020 and August 2023 at Huashan Hospital.
Imaged capillary isoelectric focusing and online mass spectrometry for milk whey protein characterization in dairy products.
Anal Biochem
April 2025
Advanced Electrophoresis Solutions Ltd., 380 Jamieson Parkway, Unit 7 and 8, ON, N3C 4N4, Canada; AES Biotech Jiaxing Ltd., No. 501 South Changsheng Road, Economic and Technological Development Zone, Jiaxing City, Zhejiang Province, PR China. Electronic address:
Characterizing major bovine milk proteins, including whey and casein, is of significant interest in the dairy industry. The diverse array of protein proteoforms can be different in terms of genetic variation, breed ways, lactation stage, and animal nutritional status. Current routine methods for bovine milk protein profiling are typically based on immunological techniques, infrared spectroscopy, slab gel isoelectric focusing, capillary electrophoresis, and high-performance liquid chromatography.
View Article and Find Full Text PDFTechnical considerations regarding saliva sample collection to achieve comparable protein identification and detection via one- and two-dimensional gel electrophoresis among humans.
Heliyon
December 2024
Department of Biochemistry and Medical Chemistry, University of Pecs, Medical School, Pecs, Hungary.
Background And Aims: Recently, demands towards identifying various molecules in support of stress detection and potential clinical utilization are dramatically increasing. Moreover, the accuracy with which researchers quantify these informative molecules is now far more improved when compared to the past. As RNA or protein markers are conventionally detected via repeated invasive procedures from blood, it is critical to develop secure technologies to obtain the desired information via less stressful methodologies, such as saliva collection.
View Article and Find Full Text PDFEvaluation of ST-segment and T-wave changes associated with NSTE-ACS in patients with RBBB: a nested case-control study design.
Intern Emerg Med
January 2025
Faculty of Medicine, Department of Emergency Medicine, Akdeniz University, Antalya, Turkey.
Patients presenting with suspected acute coronary syndrome (ACS) in the emergency department (ED) require rapid and accurate electrocardiographic (ECG) evaluation. This study aims to assess conventional ECG markers for diagnosing non-ST-elevation ACS (NSTE-ACS) in patients with chest discomfort and right bundle branch block (RBBB). A nested case-control design was employed to compare patients with RBBB admitted to the ED for suspected cardiac ischemia, focusing on those who developed NSTE-ACS versus those who did not.
View Article and Find Full Text PDFIsoelectric Focusing Fractionation Method for Signal Enhancement in Detection of Inactivated Biological Agents Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.
Electrophoresis
January 2025
National Institute for Nuclear, Chemical and Biological Protection, Kamenna, Czech Republic.
Timely identification of highly pathogenic bacteria is crucial for efficient mitigation of the connected harmful health effects. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of intact cells enables fast identification of the microorganisms based on their mass spectrometry protein fingerprint profiles. However, the MALDI-TOF MS examination must be preceded by a time-demanding cultivation of the native bacteria to isolate representative cell samples to obtain indicative fingerprints.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!