Enzyme Linked Immunosorbent Assay ELISA Test

3 min read

Enzyme Linked Immunosorbent Assay (ELISA)

Test Catageory
فئة التحليل

1. Immunolohy & Serology

Refrence Books
كتب مراجع

1. ELISA Guide Book

Skills Required
المهارات المطلوبة

1. Pipeting
2. Math
3. Attention

Video Resources
مصادر فيديو

Resource 1
Resourse عربي
Resource 3

INTRODUCTION

The enzyme-linked immunosorbent assay (ELISA) is an immunological assay commonly used to measure antibodies, antigens, proteins and glycoproteins in biological sample .
Based on how the antigen and antibodies bindThere are three types of ELISA tests .
1. Indirect ELISA: In Indirect ELISA, the antigen is attached to the test plate (kNOWN aNTIGEN)
2. Sandwich ELISA. In Sandwich ELISA, an antibody is attached to the test plate. (kNOWN Antibody)In
3. Competitive ELISA, a mixture of antigen and antibody is added to a test plate that already has antigen attached.

1. Indirect ELISA
Indirect ELISA is a test that detects antibodies in a sample. The test plate is coated with the antigen. A sample containing antibodies is added to the plate and any antibodies that bind to the antigen are detected using a secondary antibody with an attached enzyme. The secondary antibody binds to the primary antibody. Any unbound secondary antibodies are washed away. A chemical is added that reacts with the enzyme to produce a color change. The amount of color change is measured using a machine.

2. Sandwich ELISA.
Sandwich ELISA is a test that detects antigens in a sample. The test plate is coated with an antibody. A sample containing the antigen is added to the plate and any antigens that bind to the antibody are detected using a secondary antibody with an attached enzyme. The secondary antibody binds to a different part of the antigen. Any unbound secondary antibodies are washed away. A chemical is added that reacts with the enzyme to produce a color change. The amount of color change is measured using a machine.

3. Competitive ELISA
Competitive ELISA is a test that measures the amount of antigen in a sample. The test plate is coated with the antigen. Antibodies are mixed with the sample and then added to the plate. Any unbound antibodies are washed away. The more antigen in the sample, the fewer free antibodies there are to bind to the antigen on the plate. A secondary antibody with an attached enzyme is added to detect any primary antibodies on the plate. The amount of color change produced by a chemical reaction with the enzyme is measured using a machine.

Principle

ELISA is ELISA is a test that uses antibodies to detect the presence and amount of a specific substance. To make the test more accurate
and sensitive, the plate is coated with high-affinity antibodies.
it can help in the detection of an Antigen-Antibody Reaction Which is always specific for microorganism or disorder.

Equipments

1. microplate reader
2. Microplate Shaker
3. Multi channel Pipette
4. Microplates

Sample Type

Serm is usually used for the serological testing because this is where we find the antibodies that we want to detect.

Procedure

1. An antibody is attached to a solid surface, such as a polystyrene plate. The antibody can bind to bacteria, other antibodies, and hormones.
2. A mixture of antigen and antibody is added to the plate and any unbound antibodies are washed away.
3. A second antibody that binds to the first antibody is added.
4. This second antibody has an attached enzyme. Any unbound second antibodies are washed away.
5. A Substrate is added that reacts with the enzyme to produce a color change.
6. The amount of color change is measured using a machine.

Quick Notice

M. Sulieman

mohammad@mlsgaate.com

inorder to master PCR you must focus on the pipeting skills and work for enough period under supervision as long as 6 months to make sure you become professional in ELISA

crop laboratory technician examining interaction of chemicals in practical test modern lab

Enzyme Linked Immunosorbent Assay ELISA Test