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Title: ELISA TEST
Description: ELISA its Enzyme linked immuno sorbent assay,its widely used for the diagnosis of antigen or antibody present in the human body
Description: ELISA its Enzyme linked immuno sorbent assay,its widely used for the diagnosis of antigen or antibody present in the human body
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BCH 462
Enzyme-Linked Immunosorbent Assay
[ELISA]
Antigens [Ag]:
-A substance that when introduced into the body stimulates the production of an
antibody
...
They are produced by the immune system
to identify and neutralize foreign objects (antigens)
...
enzyme
Primary antibody
“antibody specified to specific antigen”
Secondary antibody
“antibody specified to primary antibody”
The enzyme linked: will convert
colorless substrate to colored product,
indicate the presence of the antibody antigen [Ab-Ag] binding complex
...
proteins (peptides, hormones) “antigens in general”
...
antibodies
...
Propose of ELISA:
-To determine the presence and the concentration of a particular Ag or Ab in a
sample
...
-In qualitative ELISA, results provide a positive or negative result for a sample
...
(obtained from serial dilutions of a
Application of ELISA:
ELISA can be used in the field of medicine, food industry and in toxicology labs to
evaluate the presence of a specific Ag or Ab in a sample
...
-Measuring hormones level
...
Polystyrene Microtiter Plate
Types of ELISA:
1
...
2
...
3
...
4
...
Direct ELISA:
-It is used to detect the presence and the concentration of specific antigen in the sample
...
Principle:
-The antigen “of interest” is adsorbed or fixed to a microtiter plate , an “enzyme is linked to an
antibody” applied to the antigen
...
By adding, the enzyme's substrate, the enzyme will convert colorless substrate to colored
product
...
-The name “direct ELISA” due to, that (the antibody linked to the enzyme) is directly bind to
the protein of interest “antigen”
...
Ag
Ag
non specific binding
...
Principle:
This method differs than direct ELISA, in that one more labeled secondary antibody
is added in the reaction
...
Then
labeled secondary antibody added that recognizes the primary antibody
...
Do not forget:
1
...
And it is the
antibody of interest in this case
...
2
...
Indirect ELISA
The name Indirect ELISA, is due to that 2ry antibody bind indirectly to the antigen
...
Principle:
The sandwich ELISA quantify antigens between two layers of antibodies (i
...
capture and detection antibody just like a sandwich)
...
The color or the signal produced as a result of addition of substrate is
proportional to antigen concentration
...
In this type of ELISA, another version of
your antigen of interest is labeled instead of the antibody
...
The color or the signal
produced as a result of addition of substrate is inversely proportional to antigens of interest
in the sample
...
Title: ELISA TEST
Description: ELISA its Enzyme linked immuno sorbent assay,its widely used for the diagnosis of antigen or antibody present in the human body
Description: ELISA its Enzyme linked immuno sorbent assay,its widely used for the diagnosis of antigen or antibody present in the human body