ELISA: How Does It Work? What are the components? What can we use it for?

Question 1

Elisa can test for __________ in a patient sample and its __________

Answer 1

Antigens, concentration

Question 2

Antibody-Antigen Chain- Why??

Answer 2

Antibody-Antigen chain is a chain created between the Antigen-Antibody. We will use Primary Antibodies to create the chain.

Question 3

Primary antibody- Why??

Answer 3

The first antibody to connect to the antigen

Question 4

Secondary antibody

Answer 4

Why?? The Secondary antibody to connect to the primary antibody. This will also have an enzyme at the end of it.

Question 5

Enzyme- Why??

Answer 5

Binds to the substrate, which will have color change if positive

Question 6

Substrate- Why??

Answer 6

The Substrate binds to the enzyme creating a bluish tint.

Question 7

Step 1

Answer 7

Label wells on strip

Question 8

Step 2

Answer 8

Add Phosphate Buffered saline to wells 2-12

Question 9

Step 3

Answer 9

Add antigen for testing

Question 10

Step 4

Answer 10

Draw out diagram, calculate dilution and concentrations

Question 11

Step 5

Answer 11

Test positive and negative control

Question 12

Step 6

Answer 12

Wait 5 minutes for protein to bind to wells

Question 13

Step 7

Answer 13

Process both strips and drain them

Question 14

Step 8

Answer 14

Fill all wells with wash buffer and drain it twice.

Question 15

Step 9

Answer 15

Transfer primary antibody into all wells and wait 5 minutes

Question 16

Step 10

Answer 16

Wait 5 minutes, then wash with buffer twice

Question 17

Step 11

Answer 17

Transfer secondary antibody into all wells

Question 18

Step 12

Answer 18

Wait 5 minutes, then wash with buffer 3 times

Question 19

Step 13

Answer 19

Transfer enzyme substrate into all wells

Question 20

Step 14

Answer 20

Wait 5 minutes

Question 21

Step 15

Answer 21

Record results and interpretations