How to set up a CIEF experiment This presentation is intended for new capillary electrophoresis users who are part of the Biomolecular Interaction Technologies.

Slides:

Advertisements

Similar presentations

Separation and Purification of Macromolecules. Why? Molecules each have specific features: Mass Charge Hyrdopathicity.

Advertisements

Isofocusing Chromatography Prepared by- Shirin Akhter.

Separation of proteins in the 1 st dimension Dr Kevin Mills Institute of Child Health, UCL, London.

9 x9 81 4/12/2015 Know Your Facts!. 9 x2 18 4/12/2015 Know Your Facts!

Capillary Electrophoresis and Capillary Electrochromatography

Protein gel electrophoresis

Electrophoretic Mobility and Electrophoresis (24.10) Electrical force is another way we can cause macromolecules to move – Macromolecules tend to have.

Protein Purification Molecular weight Charge Solubility Affinity.

PHY132 Introduction to Physics II Class 10 – Outline:

IgG IEF Diagnosing Multiple Sclerosis. Multiple Sclerosis CNS disorder Scar formation on outside of nerve cells of brain and spinal cord Inflammation.

Isoelectric Focusing Technique combining ideas of isoelectric points and electric fields Very high resolution technique for protein 1,3.

Protein Gel Electrophoresis

Proteomics: Its Function and Methods Ryan Victor.

Copyright © 2007, Pearson Education, Inc., Publishing as Pearson Addison-Wesley. Coulomb Force The field model and the electric field Chapter 20 Electric.

SFC and SF Extraction Intermediate between HPLC and GC SF are substances above their critical pressure and temperature (critical point SFs has great solvating.

Proteomics The proteome is larger than the genome due to alternative splicing and protein modification. As we have said before we need to know All protein-protein.

Electric Forces Coulomb’s Law Group Problems 1.What is the force (magnitude and direction) on a -10  C charge located at x = 4 m from a 5  C charge.

Quality Control of Product

Protein Electrophoresis BIT 230. Electrophoresis Separate proteins based on Size (Molecular Weight - MW) SDS PAGE Isoelectric Point Isoelectric focusing.

Polyacrylamide Gel Electrophoresis

Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) Mass spectrometry for protein identification 2-Dimensional Gel Electrophoresis MALDI-TOF.

Biochemical instrumental analysis-7 Dr. Maha Al-Sedik.

Experiment Two 1. Electrophoresis of serum protein (CAME) 2. Assay of casein isoelectric point ( precipitation method)

Advanced Electrophoresis Solutions Ltd

Buffers and the Henderson-Hasselbalch Equation -many biological processes generate or use H + - the pH of the medium would change dramatically if it were.

Chapter Five Protein Purification and Characterization Techniques

Electrophoresis PAGE Dr Gihan Gawish.

Analysis of Proteins and Peptides Amino acid composition Molecular weight Isoelectric point Subunit structure Prosthetic groups Solubility Biological activity.

Charging Neutral Objects Electric charge can be influenced to move by the presence of other charges (same or different) To charge a neutral object, electrons.

Advanced Analytical Chemistry – CHM 6157® Y. CAIFlorida International University Updated on 10/17/2006Chapter 9Capillary Electrophoresis Chapter 9Capillary.

Simultaneous determination of inorganic anions and cations by capillary electrophoresis with indirect UV detection I. Haumann, J. Boden, A. Mainka, U.

Amino acids/Proteins.

Capillary Electrophoresis (CE) PHAR Lecture Objectives By the end of the lecture, students should be able to: 1.Illustrate the CE instrumental.

Created with MindGenius Business 2005® Capillary Electrophoresis Capillary Electrophoresis Advantages Only needs nL sample High speed and resolution, virtually.

Isoelectric Focusing Fundamental of Bioprocess Engineering Laboratory

ANALYTICAL CHEMISTRY CHEM 3811 CHAPTER 23 DR. AUGUSTINE OFORI AGYEMAN Assistant professor of chemistry Department of natural sciences Clayton state university.

Electrophoresis: It is the movement of charged molecules in an electrical field towards the oppositely charged electrode. It is used for separation of.

Lab.3 Gel electrophoresis

General, Organic, and Biological Chemistry Copyright © 2010 Pearson Education, Inc.1 Chapter 19 Amino Acids and Proteins 19.2 Amino Acids as Zwitterions.

Keystone Review Passive Transport 1.A – ATP is a source of energy, and facilitated diffusion does not require the input of energy. 2.B – Facilitated diffusion.

Biotechnology. Gel Electrophoresis A technique that separates macromolecules on the basis of their rate of movement through a gel under the influence.

Diprotic and Polyprotic Acids: Two or more dissociable protons Consider the carbonic acid equilibrium.

KAU-Faculty of Science- Biochemistry department Analytical biochemistry lab (Bioc 343) 2012 T.A Nouf Alshareef

Welcome To my presentation.

Cell Transport. Diffusion Migration of substances from regions where their concentration is high to regions where their concentration is low. Diffusion.

Tymoczko • Berg • Stryer © 2015 W. H. Freeman and Company

Amine R group Alpha Carbon Carboxylic Acid. Nonpolar side chains.

Protein Electrophoresis & Western Blotting

Capillary electrophoresis. Principle of electrophoresis Electrophoresis is the process of moving charged molecule in solution by applying an electric.

Quality Control of Product

ELECTROPHORETIC METHODS

Tutorial 11 Modes and Methods in Capillary Electrophoresis

Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) Mass spectrometry for protein identification 2-Dimensional Gel Electrophoresis MALDI-TOF.

Amino Acids, Peptides, and Proteins

ISOELECTRIC FOCUSING Electrophoretic method that separates proteins according to the iso-electric points Is ideal for seperation of amphoteric substances.

Lecture 2 Techniques in proteomics By Ms. Shumaila Azam

“Salad” Notes Movement of molecules in and out of the cell

Separation techniques

BIOLOGY Unit 2 Notes: Cell Membrane Transport

Forging the Innovation Generation

Jefferson Method Example

Different applications of protein electrophorasis

Diagnosing Multiple Sclerosis

What is Diffusion? What does it mean to diffuse?

Capillary Electrophoresis and Capillary Electrochromatography

DNA Agarose Gel Electrophoresis

Experiment 3 Separation of proteins by Anion exchange chromatography

Víctor Rodríguez-Santiago EGEE 520

Presentation transcript:

How to set up a CIEF experiment This presentation is intended for new capillary electrophoresis users who are part of the Biomolecular Interaction Technologies Center (BITC).

Background Capillary isoelectric focusing is based on the fact that charged molecules will cease to migrate once there charge has been reduced to zero. During a CIEF experiment, ampholytes are used to produce a pH gradient across a given length of the capillary. Sample molecules then migrate under the influence of an externally applied field until they reach the point in the gradient at which the pH equals their pI.

Materials Needed: Preparing the capillary:(test to see if CE.rm video working)