Questions: Place the following components of an indirect ELISA in the correct order in which they would be added during the test procedure: 1. Patient Sample 2. Second Antibody 3. Antigen a. Substrate (A) 1,2,3,4 (B) 1,3,2,4 (C) 1,3,4,2 (D) 3,1,4,2 (E) 3,1,2,4

Place the following components of an indirect ELISA in the correct order in which they would be added during the test procedure: 1. Patient Sample 2. Second Antibody 3. Antigen a. Substrate (A) 1,2,3,4 (B) 1,3,2,4 (C) 1,3,4,2 (D) 3,1,4,2 (E) 3,1,2,4
Transcript text: Place the following components of an indirect ELISA in the correct order in which they would be added during the test procedure: 1. Patient Sample 2. Second Antibody 3. Antigen a. Substrate (A) $1,2,3,4$ (B) $1,3,2,4$ (C) $1,3,4,2$ (D) $3,1,4,2$ (E) $3,1,2,4$
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Solution

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The answer is (E): 3, 1, 2, 4

Explanation for each option:

(A) $1,2,3,4$: This order is incorrect because the antigen should be added first to coat the wells, not the patient sample.

(B) $1,3,2,4$: This order is incorrect because the antigen should be added first, not the patient sample.

(C) $1,3,4,2$: This order is incorrect because the antigen should be added first, and the substrate should be added last, not before the second antibody.

(D) $3,1,4,2$: This order is incorrect because the substrate should be added last, not before the second antibody.

(E) $3,1,2,4$: This is the correct order for an indirect ELISA. The antigen is added first to coat the wells. Then, the patient sample, which may contain antibodies specific to the antigen, is added. Next, a second antibody that binds to the patient antibodies is added. Finally, the substrate is added to produce a detectable signal.

Summary: In an indirect ELISA, the correct order of adding components is: Antigen, Patient Sample, Second Antibody, and Substrate.

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