INDIRECT ELISA IN DEPTH

Question 1

a) ___

The indirect ELISA method begins with a SAMPLE of known __ being bound to the WELLS of a ___

Answer 1

Binding Known Antigen

Antigen
Microtiter plate

Question 2

b) ___

The other UNOCCUPIED CELLS in each well are bound by a concentrated solution of ___, like ___ or ___

Answer 2

Blocking

non-interacting protein
Casein
Bovine Serum Albumin

Question 3

WHY are unoccupied sites in each well bound by a solution?

Answer 3

To BLOCK or PREVENT other proteins in the test sample from ADHERING

Question 4

c) ____

Rinse to remove any unbound ___ and ___.

Answer 4

Washing (1)

Antigen

Non-interacting protein

Question 5

d) ____

The test sample of SERUM containing the ___ is added to each well.

Examples: (3)

Answer 5

Adding Test Sample Primary Antibody

HIV antibodies
Rabies antibodies
Hepatitis B antibodies

Question 6

e) ___

Rinse to remove any ___ that did not BIND to the known ___

Answer 6

Washing(2)

Antibodies
Antigen

Question 7

f) ___

An ___ is added next to BIND to the ____.

The enzyme on (answer above) are PROTEINS, such as ___ or ____

Answer 7

Adding Enzyme-linked Secondary Antibody

Enzyme-linked secondary antibody
Test Sample

Horse radish peroxidase
Alkasline phosphatase

Question 8

g) ___

Rinse to remove any ___ that did not bind to the ___.

Answer 8

Washing (3)

Secondary antibodies
Primary antibodies

Question 9

h) ___

A ____ is then applied which is converted by the ___ to give a ___ or ___ or ___.

Answer 9

Adding substrate

Substrate
Enzyme

Color
Fluorescence
Electrochemical signal

Question 10

i) ___

By using a ___, ___, or ___, the Results can be recorded. The amount of ___ is proportional to the amount of __ bound to the ___ on the bottom of the wells.

Answer 10

Reading Results

Spectrophotometer
Spectrofluorometer
Electrochemical device

Color
Primary Antibodies
Antigen Proteins