91AV

ELISA,ELECTROPHORENSIS(SDS &NATIVE PAGE),DNA/RNA AND WESTERN BLOTTING

DATE 4^th – 8^th MARCH 2024 (5 Days)
DEADLINE FOR REGISTRATION : 23^rd FEBRUARY 2024
COST :Ksh.92800 or USD 928.00

COURSE OVERVIEW
Chrom Africa offers an outstanding platform for training on Polymerase Chain Reaction- Enzymes-linked Immunosorbent assay(PCR-ELISA) which is a commonly used analytical and biochemistry assay and or diagnostic tool in medicine,plant pathology,biotechnology and quality control check in various industries.
Delving into details the following techniques  will be covered-;

• ELISA (Enzyme-Linked Immunosorbent Assay):

ELISA is a widely used immunological method for detecting and quantifying specific proteins, peptides, antibodies, or other molecules.It involves immobilizing a target antigen or antibody on a solid surface (such as a microplate), followed by the addition of a specific enzyme-linked antibody.
The enzyme-substrate reaction produces a measurable signal (usually color change) that indicates the presence or concentration of the target molecule.

• Electrophoresis:

Electrophoresis is a technique used to separate charged molecules (such as proteins, nucleic acids, or carbohydrates) based on their size and charge.
There are two main types:

1. SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis):

Used for protein separation.
Proteins are denatured and coated with SDS, which imparts a uniform negative charge.
They migrate through a gel matrix based on size.

       2. Native PAGE (Non-denaturing Polyacrylamide Gel Electrophoresis):
Used for native protein analysis (maintains protein structure).
No SDS is used, and proteins retain their native charge.
Migration depends on both size and charge.

• DNA and RNA:

DNA (Deoxyribonucleic Acid):
The genetic material in all living organisms.
Composed of nucleotides (adenine, thymine, cytosine, and guanine) arranged in a double-stranded helix.
Stores genetic information and serves as a template for RNA synthesis.

RNA (Ribonucleic Acid):
Single-stranded nucleic acid.
Types include messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA).
Involved in protein synthesis, gene regulation, and other cellular processes.

• Western Blotting (Immunoblotting):

A technique to detect and analyze specific proteins in a complex mixture.
Steps:

Protein Separation:
Proteins are separated by SDS-PAGE or other electrophoresis methods.
Transfer to Membrane:
Proteins are transferred from the gel to a membrane (usually nitrocellulose or PVDF).
Blocking:
The membrane is blocked to prevent nonspecific binding.
Primary Antibody Incubation:
Specific primary antibodies bind to the target protein.
Secondary Antibody Incubation:
Secondary antibodies (conjugated to enzymes or fluorophores) bind to the primary antibodies.
Detection:
Enzymatic or fluorescent substrates generate a signal, revealing the presence of the target protein13.

SUITABILITY
This course is suitable for researchers, scientists, laboratory analysts, graduate students and postgraduate students who have a background in molecular biology/biotechnology,analytical chemistry or biochemistry, who are interested in learning more about the practical aspects of the course.