ELISAvirology

Hamid Afkhami

EL Enzyme-LinkedEnzyme catalysis the substrate and produce colored species.

IS ImmunoSorbentAntigen or antibody are sticked in wells

About ELISA

Types of the ELISA1. Direct2. Indirect3. Sandwich4. competitive

Direct ELISASTEPS:

Antigen stick to well with charge + casein or albumin(BSA).

Add sample to well Wash Add substrate acid measure OD

Advantages Quick Eliminate Cross-

reactivity of secondary Ab

Disadvantages the signal is less

amplified Labeling has destructive

effects on ab and consuming time.

Direct ELISA

STEPS: Antigen stick to well with

charge + casein or albumin(BSA).

Add sample + Wash well Add detector ab + wash Add substrate and

measure OD

Indirect ELISA

Advantages: High sensitivity Flexibility This format of indirect ELISA is

suitable for determining total antibody level in samples

Cost saving Several Epitopes for secondary

Ab and signal amplification

Indirect ELISA

Antigen detection STEPS:

Stick ab to well + BSA Add sample (ag) + wash Add detector ab + wash Add substrate + measure OD

Advantages High specificity Suitable for complex samples

SANDWICH ELISAcontain at least two antigenic

COMPETITIVE ELISA

COMPETITIVE ELISA

Advantages Suitable for complex samples Flexibility and sensitivity

COMPETITIVE ELISA

http://www.elisa-antibody.com/ELISA-Introduction https://www.bio-rad-antibodies.com https://www.youtube.com http://arian-andish.com https://en.wikipedia.org/wiki/ELISA

Sources

Have a nice day