Research Method

Step 1

Labeled hormone

(unbound)

Labeled antibody

Step 2

Unlabeled antibody

Step 3

Step 1

Labeled hormone

(unbound)

Labeled antibody

Step 2

Add a saturating amount of labeled hormone to a stationary antibody.

Wash out unbound hormone and measure amount of labeled hormone bound to antibodies.

O Unlabeled hormone

Add a known quantity of unlabeled hormone, which will displace some of the labeled hormone bound to the antibodies.

Wash out unbound hormone and measure amount of labeled hormone bound to antibodies.

Add a saturating amount of labeled hormone to a stationary antibody.

Wash out unbound hormone and measure amount of labeled hormone bound to antibodies.

O Unlabeled hormone

Add a known quantity of unlabeled hormone, which will displace some of the labeled hormone bound to the antibodies.

Duplicate the conditions in step 1 and add a larger, known quantity of unlabeled hormone that will displace even more of the labeled hormone.

Wash out unbound hormone and measure amount of labeled hormone bound to antibodies.

Wash out unbound hormone and measure amount of labeled hormone bound to antibodies.

O Unlabeled hormone

Duplicate the conditions in step 1 and add a larger, known quantity of unlabeled hormone that will displace even more of the labeled hormone.

Wash out unbound hormone and measure amount of labeled hormone bound to antibodies.

The dose that stimulates half the maximum response is a measure of sensitivity to the hormone.

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