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ELCTROPHORESIS
by:- Naman goyal
roll no. 72
ELECTROPHORESIS
 DEFINATION: It describes migration of charged particles or
molecules under the influence of electric field.
 PURPOSE FOR CRRYING OUT ELEECTROPHORESIS
 1. To determine the number, amount and mobility of
components in given sample or to separate them.
 2. Determination of molecular weight of proteins and DNA
sequencing.
 3. To obtain information abut the electrical double layers
surrounding the particles.
 FACTORS AFFECTNG ELECTROPHORETIC MOBILITY
 1. CHARGE
 2. SIZE
 3. SHAPE
 TYPES OF ELECTOPHORESIS
 1. Zone Electrophoresis
 a) Paper Electrophoresis
 b) Gel Electrophoresis
 2. Isoelectric Focussing
 3. Immunoelectrophoresis
 PAPER ELECTROPHORESIS
 It is the form of electrophoresis that is carried out on filter paper.
This technique is useful for separation of small charged molecules
such as amino acids and small proteins.
 The serum proteins are separatd into 5 distinct bands- albumin,
α1-, α2-, β- and γ-glbulins.
 GEL ELECTROPHORESIS
 It is a technique used for the separation of DNA, RNA, or protein
molecules according to their size and charge using an electric current
applied to the gel matrix.
 The serum proteins can be separated to about 15 distinct bands.
 Types of the gel:
 1. Agarose gel
 2. Polyacrylamide gel
 3. Sodium dodecyl sulphate (SDS)
Paper Electrophoresis Gel Electrophoresis
 Polyacrylamide is employed for the determination of
molecular weights of proteins in a popularly known
electrophoresis technique known as SDS-PAGE.
 ISOELECTRIC FOCUSSING
 This technique is based on the immobilization of the molecules at
isoelectric pH during electrophoresis.
 It is ideal for separation of amphoteric substances.
 It’s gels contain synthetic buffers called ampholytes that smooth the pH
gradients.
 The serum proteins can be separated to as many as 40 bands.
 IMMUNOELECTROPHORESIS
 It is atwo stage process, Electrophoresis is conducted in first stage and
immunoprecipitation using antibodies against specific proteins in the
second stage.
 The antibodies when come in contact with antigens, precipitation occurs,
resulting in the formation of precipitin bands.
Isoelectric focussing Immunoelectrophoresis

THANK YOU

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Electrophoresis

  • 2. ELECTROPHORESIS  DEFINATION: It describes migration of charged particles or molecules under the influence of electric field.  PURPOSE FOR CRRYING OUT ELEECTROPHORESIS  1. To determine the number, amount and mobility of components in given sample or to separate them.  2. Determination of molecular weight of proteins and DNA sequencing.  3. To obtain information abut the electrical double layers surrounding the particles.  FACTORS AFFECTNG ELECTROPHORETIC MOBILITY  1. CHARGE  2. SIZE  3. SHAPE
  • 3.  TYPES OF ELECTOPHORESIS  1. Zone Electrophoresis  a) Paper Electrophoresis  b) Gel Electrophoresis  2. Isoelectric Focussing  3. Immunoelectrophoresis
  • 4.  PAPER ELECTROPHORESIS  It is the form of electrophoresis that is carried out on filter paper. This technique is useful for separation of small charged molecules such as amino acids and small proteins.  The serum proteins are separatd into 5 distinct bands- albumin, α1-, α2-, β- and γ-glbulins.  GEL ELECTROPHORESIS  It is a technique used for the separation of DNA, RNA, or protein molecules according to their size and charge using an electric current applied to the gel matrix.  The serum proteins can be separated to about 15 distinct bands.  Types of the gel:  1. Agarose gel  2. Polyacrylamide gel  3. Sodium dodecyl sulphate (SDS)
  • 5. Paper Electrophoresis Gel Electrophoresis
  • 6.  Polyacrylamide is employed for the determination of molecular weights of proteins in a popularly known electrophoresis technique known as SDS-PAGE.  ISOELECTRIC FOCUSSING  This technique is based on the immobilization of the molecules at isoelectric pH during electrophoresis.  It is ideal for separation of amphoteric substances.  It’s gels contain synthetic buffers called ampholytes that smooth the pH gradients.  The serum proteins can be separated to as many as 40 bands.  IMMUNOELECTROPHORESIS  It is atwo stage process, Electrophoresis is conducted in first stage and immunoprecipitation using antibodies against specific proteins in the second stage.  The antibodies when come in contact with antigens, precipitation occurs, resulting in the formation of precipitin bands.