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picture1_Responsibility Ppt 77498 | 5991 8063en Agilent Ce Theory


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File: Responsibility Ppt 77498 | 5991 8063en Agilent Ce Theory
agilent technologies is committed to the educational community and is willing to provide access to company owned material contained herein this slide set is created by agilent technologies the usage ...

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                                            Agilent Technologies is committed to the 
                                            educational community and is willing to provide 
                                            access to company-owned material contained 
                                            herein.
                                            This slide set is created by Agilent Technologies. The usage of the slides is 
                                            limited to teaching purpose only.
                                            These materials and the information contained herein are accepted “as is” and 
                                            Agilent makes no representations or warranties of any kind with respect to the 
                                            materials and disclaims any responsibility for them as may be used or 
                                            reproduced by you.  Agilent will not be liable for any damages resulting from or 
                                            in connection with your use, copying or disclosure of the materials contained 
                                            herein.  You agree to indemnify and hold Agilent harmless for any claims 
                                            incurred by Agilent as a result of your use or reproduction of these materials.
                                            In case pictures, sketches or drawings should be used for any other purpose 
                                            please contact Agilent Technologies a priori.
   For teaching purpose only
   9/3/22  5991-8063EN
   © Agilent Technologies, Inc. 2016
   2
   Introduction
   Electrophoresis has been defined as the differential movement of charged 
   species (ions) by attraction or repulsion in an electric field. 
   Electrophoresis as a separation technique was introduced by Arne Tiselius 
   (Sweden, 1902-1971) in 1937. Placing protein mixtures between buffer 
   solutions in a tube and applying an electric field, he found that sample 
   components migrated in a direction and at a rate determined by their charge 
   and mobility. 
   For his work in separation science Tiselius was awarded a the 
   Nobel Prize for Chemistry (1948).
   For teaching purpose only. 
   9/3/22  5991-8063EN
   © Agilent Technologies, Inc. 2016
   3
   Table of Contents (ToC)
   Introduction
   • How does it work?
   Principles
   • Electrophoresis - Ion velocity
   • Electrophoresis - Forces
   • Electrophoresis - Mobility
   • Electroosmotic flow
   • Mobility and migration time
   • Resolution
   Comparison of techniques
   • Chromatography vs Electrophoresis
   Characteristics of capillary electrophoresis
   Further Information
   For teaching purpose only. 
   9/3/22  5991-8063EN
   © Agilent Technologies, Inc. 2016
   4
    Introduction
    How does it work?
    Electro (electric) + Phoresis (migration) 
                                                                                    Negative electrode
    Simplified process:
    •    Samples are loaded into wells and                                             n
                                                                                       o
                                                                                    &  i
         charged molecules start migrating                                             t
                                                                                    d  c
                                                                                    l  e
                                                                                    e  r
                                                                                    i  i
         through the gel when voltage is                                            f  d
                                                                                        
                                                                                    c  n
                                                                                    i
                                                                                    r  o
                                                                                    t  i
         applied.                                                                   c  t
                                                                                    e  a
                                                                                    l  r
                                                                                    E  g
                                                                                       i
    •    Molecules with smaller molecular                                              m
         weight move faster then molecuels 
         with higher molecular weight. 
                                                                                    Positive electrode
    ToC
    For teaching purpose only. 
    9/3/22  5991-8063EN
    © Agilent Technologies, Inc. 2016
    5
    Introduction
    How does it work?
      Separation efficiency in free solution, 
      as performed by Tiselius, was limited                                     (a)                                            (b)
      by thermal diffusion and convection.
      For this reason, electrophoresis 
      traditionally has been performed in 
      anti-convective media, such as 
      polyacrylamide or agarose gels. 
      Primary application area:
      •   Size-dependent separation of 
          biological macromolecules such as 
          nucleic acids (RNA/DNA) and 
          proteins.                                                           Comparison of gel used for slab gel electrophoresis (a) 
                                                                              and capillary for Capillary electrophoresis (b).
    ToC
    For teaching purpose only. 
    9/3/22  5991-8063EN
    © Agilent Technologies, Inc. 2016
    6
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...Agilent technologies is committed to the educational community and willing provide access company owned material contained herein this slide set created by usage of slides limited teaching purpose only these materials information are accepted as makes no representations or warranties any kind with respect disclaims responsibility for them may be used reproduced you will not liable damages resulting from in connection your use copying disclosure agree indemnify hold harmless claims incurred a result reproduction case pictures sketches drawings should other please contact priori en inc introduction electrophoresis has been defined differential movement charged species ions attraction repulsion an electric field separation technique was introduced arne tiselius sweden placing protein mixtures between buffer solutions tube applying he found that sample components migrated direction at rate determined their charge mobility his work science awarded nobel prize chemistry table contents toc ho...

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