ELISA and Western blot| 5 Similarities & 12 Differences

ELISA (ENZYME LINKED IMMUNO SORBITANT ASSAY) and Western blot areĀ  both immuno assays.

They rely on use of antigens and antibodies in the reaction.

Both these experiments are analytical in nature and used to detect biomolecules like proteins, enzymes or other pathogens.

They are widely used in molecular biology, medical research and also in health care.

ELISA is a quantitative technique which is rapid and relies heavily on production of colors.

While western blot is semi-quantitative technique which relies on production of separated bands of biomolecules. It employs gel electrophoresis to separate the proteins.

Elisa and Western blot

Both these techniques are based on immunology principle but, they have different procedures and applications. So we will see them as similarities and differences.

ELISA vs Western Blot similarities.

Since they follow same principle there are few similarities in between them

Elisa and Western Blot Similarities

SL.NOELISAWestern Blot
PrincipleBased on Immunological assayImmunological assay
Method of assayReaction between an antigen and antibody forming a complexAntigen antibody reactions to form a complex
Type of sampleBiological samplesBiological samples only.
Compounds than can be analyzedProteins, enzymes, hormones, steroidsEnzymes, proteins, hormones.
Elisa and Western blot

Credit: Bio-rad

But they are many differences between these two techniques. These differences help the researcher decide which of the one of the two to take up.

Elisa vs Western Blot Differences

SL.NoELISAWestern Blotting
DetectsAntibody or antigen in the tissueMostly the antigen in the tissue
Time for experimentVery fast like in couple of minutesVery time taking like 2 to 3 days.
Number of samples per experimentMany sample can be analyzed in a single run like 96 samples per turn using plate readersOnly limited samples like 7 samples per experiment is feasible.
Cost of experimentComparatively less expensive.Very expensive as it requires many basic chemical, gels and even blotting paper.
Accuracy of detectionThe sample detection is very accurate and even the quantity can be clearly known.Though accurate, quantity can not be clearly known.
Technique for operationIt is very easy and even a new experimenter can do with little guidance.Very complicated and requires practice and standardization of whole procedure.
Sensitivity is enhanced byWashing and removal of unbound proteins.Here other interfering proteins are removed by gel electrophoresis.
Types of experimentsELISA is of different types like direct, indirect, sandwich type.It is of different types like southern blot, western and even northern blot.
Pictorial demonstrationELISA is just based on readings by electronic instruments. So it gives only result but no pictureThis is based on physical appearance so enables to take photos and depict the result in presentations and publications.
ApplicationMostly in health care for diagnosis and also in research.Mostly in research but less preferred in health care due to time span.
Age of the techniqueIt is newer techniqueOlder technique

It appears that those who need a pictorial demonstration for publications and presentations for western blot technique. While, those who need just an accurate result like in health care go for ELISA.

In health care ELISA is used to identify many diseases like AIDS, Hepatitis and other infections by just taking serum sample. It is also widely used in research as it is less time consuming and easy to perform.

On other hand, Western blotting test may be used as a confirmatory test for HIV after ELISA to avoid any false positives.

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