Capillary electrophoresis is a separation technique where electrophoresis is performed inside narrow capillaries. Charged molecules or ions migrate through the capillary under the influence of an electric field. The rate of migration depends on factors like the molecule's net charge, size, shape, and the electric field strength. There are two main types - capillary zone electrophoresis, which separates analytes in buffer alone, and capillary gel electrophoresis, which uses a gel matrix to separate based on size. Capillary electrophoresis has applications in fields like pharmaceuticals, forensics, foods, and biosciences for analyzing substances like DNA, proteins, metals, and organic compounds.
Introduction
History
Elecrophoresis
Principle
Types of electrophoresis
Application
Conclusion
Reference
When a potential difference is applied between the two electrodes in a colloidal solution, It has been observed that the colloidal particles are carried to either the positive or negative electrode.
In other words , they behave as if they are electrically charged w.r.t. the dispersion medium. This phenomenon is known as electrophoresis.
Many important biological molecules, such as amino acids, peptides, proteins, nucleotides and nucleic acids, possess ionisable groups and, therefore, at any given pH, exist in solution as electrically charged species either as cations or anions.
Under the influence of an electric field these charged particles will migrate either to the cathode or to the anode, depending on the nature of their net charge.
The technique of paper electrophoresis is simple and inexpensive and requires only micro quantities of plasma for separation.
The support medium is a filter paper
The electrophoresis apparatus in its simplest form consists of two troughs to contain buffer solution, through which electric current is passed.
Frequently used in isolating proteins, amino acids and oligopeptides.
Electrophoresis principle and types by Dr. Anurag YadavDr Anurag Yadav
Â
the general principle on how the electrophoresis performs.
the different types of electrophoresis and the mechanism of separation based on different character of the medium and type of electrophoresis.
Sepration of molecules on the basis of applied Electric Field
Categorized into 1) Zone Electrophoresis 2) Moving Boundary Electrophoresis
We can seprate macromolecules (DNA , RNA, PROTEINS )on the basis of their charge, size shape & molecular weight
In this slide contains types, working principle, factors affecting, advantage and disadvantage of paper electrophoresis.
Presented by: G.Sai Swetha. (Department of pharmacology),
RIPER, anantapur.
Introduction
History
Elecrophoresis
Principle
Types of electrophoresis
Application
Conclusion
Reference
When a potential difference is applied between the two electrodes in a colloidal solution, It has been observed that the colloidal particles are carried to either the positive or negative electrode.
In other words , they behave as if they are electrically charged w.r.t. the dispersion medium. This phenomenon is known as electrophoresis.
Many important biological molecules, such as amino acids, peptides, proteins, nucleotides and nucleic acids, possess ionisable groups and, therefore, at any given pH, exist in solution as electrically charged species either as cations or anions.
Under the influence of an electric field these charged particles will migrate either to the cathode or to the anode, depending on the nature of their net charge.
The technique of paper electrophoresis is simple and inexpensive and requires only micro quantities of plasma for separation.
The support medium is a filter paper
The electrophoresis apparatus in its simplest form consists of two troughs to contain buffer solution, through which electric current is passed.
Frequently used in isolating proteins, amino acids and oligopeptides.
Electrophoresis principle and types by Dr. Anurag YadavDr Anurag Yadav
Â
the general principle on how the electrophoresis performs.
the different types of electrophoresis and the mechanism of separation based on different character of the medium and type of electrophoresis.
Sepration of molecules on the basis of applied Electric Field
Categorized into 1) Zone Electrophoresis 2) Moving Boundary Electrophoresis
We can seprate macromolecules (DNA , RNA, PROTEINS )on the basis of their charge, size shape & molecular weight
In this slide contains types, working principle, factors affecting, advantage and disadvantage of paper electrophoresis.
Presented by: G.Sai Swetha. (Department of pharmacology),
RIPER, anantapur.
Electrophoresis is the movement of charged particles through an electrode when subjected to an electric Field
Cations move towards cathode
Anions move towards anode
By this technique solutes are separated by their different rates of travel through an electric field.
Commonly used in biological analysis, particularly in the separations of proteins, peptides and nucleic acids
Isoelectric focusing electrophoresis- Principle , procedure and applicationsJaskiranKaur72
Â
IEF separates amphoteric compounds, such as proteins, with increased resolution in a medium possessing a stable pH gradient. The protein becomes âfocusedâ at a point on the gel as it migrates to a zone where the pH of the gel matches the protein's pI. At this point, the charge of the protein becomes zero and its migration ceases.
Separation is brought about through molecular sieving technique, based on the molecular size of the substances. Gel material acts as a "molecular sieveâ.
Gel is a colloid in a solid form (99% is water).
It is important that the support media is electrically neutral.
Different types of gels which can be used are; Agar and Agarose gel, Starch, Sephadex, Polyacrylamide gels.
Introduction, Principle, Instrumentation and Applications of SDS-PAGEMohammed Mubeen
Â
The following presentation contains helpful information regarding SDS-PAGE, including the history, introduction, principle, instrumentation, advantages and applications of SDS-PAGE.
Electrophoresis:
Electrophoresis is separation technique based on movement of charge particle in an electric field.
Movement of charge particles can be determined by following formula--
V= Eq/f
Where,
V= Velocity of the charged particle;
E= electric field of the molecule;
q= Net charge of the molecule; and
f= Frictional co-efficient of the molecule
Types of electrophoresis:
1. Agarose gel electrophoresis ;
2. Poly-acryl amide gel electrophoresis [PAGE];
3. Sodium do-decyl sulphate Poly- acrylamide gel electrophoresis [SDS-PAGE] ;
4. Two dimensional âPoly-acrylamide gel electrophoresis [2D-PAGE];
5. Pulse field gel electrophoresis [PFGE];
6. Capillary gel electrophoresis [CGE]; and
7. Disc electrophoresis for Protein.
Application of electrophoresis:
1. Estimation of the DNA molecule.[ Agarose , PAGE ]
2. Analysis of PCR product. [ Agarose ]
3. Separation of restricted genomic DNA and RNA. [Agarose and PAGE respectively]
4. Conformation of newly isolated DNA .[Agarose]
5. Separation of most small fragments of DNA. [PAGE]
6. In forensic science.[Agarose , PAGE, SDS-PAGE, 2D PAGE ,Capillary gel electrophoresis , PFGE]
8. In determining molecular wt. of protein.[SDS-PAGE].etc
Electrophoresis is the movement of charged particles through an electrode when subjected to an electric Field
Cations move towards cathode
Anions move towards anode
By this technique solutes are separated by their different rates of travel through an electric field.
Commonly used in biological analysis, particularly in the separations of proteins, peptides and nucleic acids
Isoelectric focusing electrophoresis- Principle , procedure and applicationsJaskiranKaur72
Â
IEF separates amphoteric compounds, such as proteins, with increased resolution in a medium possessing a stable pH gradient. The protein becomes âfocusedâ at a point on the gel as it migrates to a zone where the pH of the gel matches the protein's pI. At this point, the charge of the protein becomes zero and its migration ceases.
Separation is brought about through molecular sieving technique, based on the molecular size of the substances. Gel material acts as a "molecular sieveâ.
Gel is a colloid in a solid form (99% is water).
It is important that the support media is electrically neutral.
Different types of gels which can be used are; Agar and Agarose gel, Starch, Sephadex, Polyacrylamide gels.
Introduction, Principle, Instrumentation and Applications of SDS-PAGEMohammed Mubeen
Â
The following presentation contains helpful information regarding SDS-PAGE, including the history, introduction, principle, instrumentation, advantages and applications of SDS-PAGE.
Electrophoresis:
Electrophoresis is separation technique based on movement of charge particle in an electric field.
Movement of charge particles can be determined by following formula--
V= Eq/f
Where,
V= Velocity of the charged particle;
E= electric field of the molecule;
q= Net charge of the molecule; and
f= Frictional co-efficient of the molecule
Types of electrophoresis:
1. Agarose gel electrophoresis ;
2. Poly-acryl amide gel electrophoresis [PAGE];
3. Sodium do-decyl sulphate Poly- acrylamide gel electrophoresis [SDS-PAGE] ;
4. Two dimensional âPoly-acrylamide gel electrophoresis [2D-PAGE];
5. Pulse field gel electrophoresis [PFGE];
6. Capillary gel electrophoresis [CGE]; and
7. Disc electrophoresis for Protein.
Application of electrophoresis:
1. Estimation of the DNA molecule.[ Agarose , PAGE ]
2. Analysis of PCR product. [ Agarose ]
3. Separation of restricted genomic DNA and RNA. [Agarose and PAGE respectively]
4. Conformation of newly isolated DNA .[Agarose]
5. Separation of most small fragments of DNA. [PAGE]
6. In forensic science.[Agarose , PAGE, SDS-PAGE, 2D PAGE ,Capillary gel electrophoresis , PFGE]
8. In determining molecular wt. of protein.[SDS-PAGE].etc
electrophoresis: movement of charge particles in a gel under the influence of an electric field, principle, factors, apparatus, types , application, advantage and disadvantage.
Electrophoresis is an electrokinetic process which separates charged particles in a fluid using a field of electrical charge. It is most often used in life sciences to separate protein molecules or DNA and can be achieved through several different procedures depending on the type and size of the molecules. The procedures differ in some ways but all need a source for the electrical charge, a support medium and a buffer solution. Electrophoresis is used in laboratories for the separation of molecules based on size, density and purity.An electric field is applied to molecules and as they are electrically charged themselves it results in a force acting upon them. The greater the charge of the molecule the greater the force applied by the electrical field and therefore the further through the support medium the molecule will move relative to its mass.
Some example applications of electrophoresis include DNA and RNA analysis as well as protein electrophoresis which is a medical procedure used to analyse and separate the molecules found in a fluid sample (most commonly blood and urine samples).Different types of gels are usually used as the support medium for electrophoresis and this may be in slab or tube form depending on which is more beneficial. Gel slabs enable many samples to be run simultaneously and so are frequently used in laboratories. However, tube gels give a better resolution of the results so are often chosen for protein electrophoresis.
Agarose gel is commonly used for electrophoresis of DNA. It has a large pore structure allowing larger molecules to move easily but it is not suitable for sequencing smaller molecules.
Polyacrylamide gel electrophoresis (PAGE) has a clearer resolution than agarose gel making it more suitable for quantitative analysis. This makes it possible to identify how proteins bind to DNA. It can also be used to develop an understanding of how bacteria is becoming resistant to antibiotics through plasmid analysis.
This presentation contain the information about gel electrophoresis method , instruments & types.
Electrophoresis is a method through biological molecules are separated by applying an electric field.
Main purpose of this method is to determine the number , amount & mobility of biological component.
There are some internal & external factors that affects the process of electrophoresis.
The bio-molecules have charge on it & when we apply an electric field , the charge particles move to the opposite cathode. In this way, charge particles are separated
There are 3 types of gels that use in this process .
In this buffers are also used which provide ions that carry a current.
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Electrophoresis , principles and application
1. ď§Principle:
Any charged ion or
molecule migrates when
placed in an electric field
the rate of migration
depend upon its net
charge size,shape and the
applied electric current.
ď§ Mechanism:
⢠Definition=âmigration of charged particles or
molecules under the influnce of electric current.â
⢠Literally= greek word means transport by
electricity.
Electrophoresis
2. Factors affecting the rate of ion
mobility
â Net charge of
molecule
â Size & shape of
molecule
â Electric field
strength
â Properties of
supporting medium
â Temperature
â pH of the buffer
3. Capillary electrophoresis:
Sample vial Destination vial
Definition;
It is a seperation
technique in which
electrophoresis is
performed in narrow bore
capillaries typically 25-
100um inner diameter.
⢠Difference from
electrophoresis;
In traditional
electrophoresis,electricall
y charged analytes move
in a conductive liquid
medium under the
influence of an electric
field,whereas cappilary
electrophoresis is
designed to separate
species based on their
charge to size ratio in the
interior of small capillary
filled with an electrolye.
4. Instrumentation
ď A fused silica capillary
ď Sample vial and destination vial
ď Two buffer reserviors
ď A high voltage power supply
ď Detector
ď Data output and handling device
6. 1-Capillary zone electrophoresis
(CZE)
PRINCIPLE âIn
CZE,analytes
are seperated
in a capillary
containing only
buffer,without
any anti
convective
mediumâ
Mechanism:
EXAMPLE:
Separation of small
ions, some proteins,
amino acids, and
carbohydrate.
7. 2-Capillary gel electrophoresis(CGE)
ď§ Principle;
âseperation takes place
inside a capillary filled
with a gel which acts
as a molecular sieveâ
Mechanism;
ď§ Gels:
ď§ linear noncrosslinked
polyacrylamide
(CH2=CH-CO-NH2) (C-
PAGE)
ď§ dextran,
ď§ agarose, and
ď§ poly(ethyleneoxide)
⢠Gel matrix viscosity, density, and
pore size are all factors in
determining the âspeedâ of
separation.
⢠Two types of separation can be done
using GE:
o Native: separation by size
and
o charge (charge/mass)
o Denaturing: separation by
size
8. Instrumentation
⢠Power supply
⢠Cooling Apparatus
⢠Electrophoresis gel
apparatus
⢠White Light/UV Light Box
⢠Digital Camera/Gel
Documentation System
⢠Working:
⢠Prepare samples
⢠Prepare gel and buffers
⢠Load samples onto gel
⢠Run gel
⢠Stain gel
⢠Interpret/analysis of gel
⢠Archive (photograph, dry
gel)
9. Application
⢠Determinatio
n of DNA
sequencesâŚ.
⢠Formation of
gel
(agarose+buff
er)
⢠Heat it and
allowed to
cool down
⢠Place the gel
in buffer
⢠Application of
samples
10.
11. Applications of capillary
electrophoresis
⢠Following are the fields in which electrophoresis play
important role:
⢠Pharmaceutics
⢠Forensic
⢠Foods
⢠Bioscience
⢠Agricultural field
⢠Pesticide analysis
⢠Surfactants analysis
⢠Transition metal analysis
⢠Organic compound analysis