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Name#Mukhdoma jaffary
roll# 23011760-016
topic# PAGE
presented to : Dr Muddassar hussain
PAGE:
Polyacrylamide gel electrophoresis (PAGE) is a technique
widely used in biochemistry, forensic chemistry,
genetics, molecular biology and biotechnology to
separate biological macromolecules, usually proteins or
nucleic acids, according to their electrophoretic
mobility.
Principle :
Protocol:
1. Sample preparation:
 Protein samples are denatured by heating them with a
detergent SDS and mercaptoethanol.
 The former binds strongly to the proteins and gives
them a high negative charge
 This helps the resolution of proteins strictly based on
their size during gel electrophoresis.
Mechanism:
2. Gel preparation:
3. Electrophoresis:
 As an electric current is applied proteins migrate
through the gel to the positive electrode as they have
a negative charge.
 Each molecule moves at a different rate based on its
molecular weight
 After a few hours, the protein molecules are all
separated by size.
4. Staining and visualization:
the gel can be stained using colored dyes such as
 Coomassie Brilliant Blue or ethidium bromide
 bromophenol blue along with the protein sample
Applications:
SDS-PAGE has many applications. It is mostly used for
following purposes:
1. Establishing protein size
2. Protein identification
3. Determining sample purity
4. Identifying disulfide bonds
5. Quantifying proteins
6. Blotting applications
PAGE.pptxr67dytcyg7tr6dtfygu7bguyfutcgvhgu7tfuyv
PAGE.pptxr67dytcyg7tr6dtfygu7bguyfutcgvhgu7tfuyv

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  • 1. Name#Mukhdoma jaffary roll# 23011760-016 topic# PAGE presented to : Dr Muddassar hussain
  • 2. PAGE: Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility.
  • 4. Protocol: 1. Sample preparation:  Protein samples are denatured by heating them with a detergent SDS and mercaptoethanol.  The former binds strongly to the proteins and gives them a high negative charge  This helps the resolution of proteins strictly based on their size during gel electrophoresis.
  • 7. 3. Electrophoresis:  As an electric current is applied proteins migrate through the gel to the positive electrode as they have a negative charge.  Each molecule moves at a different rate based on its molecular weight  After a few hours, the protein molecules are all separated by size.
  • 8. 4. Staining and visualization: the gel can be stained using colored dyes such as  Coomassie Brilliant Blue or ethidium bromide  bromophenol blue along with the protein sample
  • 9. Applications: SDS-PAGE has many applications. It is mostly used for following purposes: 1. Establishing protein size 2. Protein identification 3. Determining sample purity 4. Identifying disulfide bonds 5. Quantifying proteins 6. Blotting applications