Electrophoresis is a laboratory technique

1. Prepared by :
Mzhda Salman
Hawrin Ibrahim
Hawler medical University
College of medicine
Biochemistry
Electrophoresis

2. 1. It is the process of moving charged biomolecules
in solution by applying an electrical field across the
mixture.
2. Biomolecules moved with a speed dependent on
their charge, shape, and size and separation
occures on the basis of molecular size.
Basic principles of electrophoresis
Introduction

3. When charged molecules are placed in
an electric field, they migrate
toward either the positive (anode)
or negative (cathode) pole
according to their charge.
Factors influenced electrophoresis
1. mobility:
2. net charge of the molecule
3. size and shape
4. concentration of the molecule in
solution

4. Types of electrophoresis
1- Gel electrophoresis
2- Moving boundary or frontal electrophoresis
3- Zonal electrophoresis : this type is mach less costly also much
less material is required and many runs can be carried out at
the same time
4- Capillary Gel Electrophoresis (CGE)

5. Gel electrophoresis
 It is a technique used for the speration of Deocyribonucleic
acid,ribonucleic acid or protein molecules according to their size
and electrical charge using an electric current applied to a gel
matrix
 What is Gel?gel is a cross linked polymer whose composition and
porosity is chosen based on a specific weight and porosity of the
target molecules
 Types of Gel:
 Agarose Gel
 Polyacrylamide gel

6. Zone electrophoresis
Is the various fractions of serum proteins (albumens and globulins).
• Much simple method
• Much greater resolution
• Require small sample
• Acetate cellulose – support medium
Protein separation depends on :
• Type and number of ionizable side chains of amino acids - R.
• Size of net charge (positive or negative).
• Negatively charged proteins move towards the anode.
• Positively charged proteins move towards the cathode.

8. Importance of electrophoresis :
These days this technique is being extensively used for
1 - the separation of aminoacids and proteins.
2- this technique is of great diagnostic significance is several disease
/disorders because it has been found that the electrophoretic proteins of
human plasma may vary widely from the normal in pathological states for
example .
3-This technique can be used to identify genes in humans that have
previously been identified in other organisms.
4-It can also be used to compare the DNA found from blood or hair
samples at a crime scene with the DNA of a suspect in the crime.
Diseases diagnosed with electrophoresis
1-Sickle Cell Anemia
2-thalassemia

9. An ethidium-stained gel photographed under UV light
**Each band that you see is a collection of millions of
DNA molecules, all of the same length!!

10. Protein Fraction
Normal Curve
Albumin
Globulin Fractions
γ β α2 α1
con

11. Albumin
Globulin Fractions
γ β α2 α1
Con
Protein Fraction
1- In chronic inflammatory disease
A- Increased alpha one alpha two ,beta and gamma
B- Decrease in albumin fruction

12. 2- In multiple myeloma
increased in globulin and show M band with a light decreased
In albumin ,alpha one ,alpha two and beta fraction
Albumin
Globulin Fractions
γ β α2 α1
Con
Protein Fraction

13. 3 - in liver disease (cirrhosis )
A-Incrased in gama and beta fruction
B-Decrease in albumin and alpha one and alpha two
Albumin
Globulin Fractions
γ β α2 α1
Con
Protein Fraction

14. 4- In nephritic syndrome
a. increased in alpha two and marked decreased in
albumin
Albumin
Globulin Fractions
γ β α2 α1
Con
Protein Fraction

15. 5 - Bone cancer
decreased in globulin nearly absent
Albumin
Globulin Fractions
γ β α2 α1
Con
Protein Fraction

16. Thank
You for
listening