Normalization methods for G-protein coupled receptor membrane array
First Claim
Patent Images
1. A reference prelabeled G-protein coupled receptor membrane array comprising a plurality of assayable microspots, wherein one or more of the plurality of the microspots comprise:
- a membrane comprising (i) a target G-protein coupled receptor embedded the membrane and (ii) a membrane component; and
a labeled agent bound to the membrane component.
1 Assignment
0 Petitions
Accused Products
Abstract
Reference membrane components are either pre-labeled or labeled during assays for purposes of normalizing signals associated with binding or functional assays employing G-protein coupled receptor microarrays. A reference component may be included in a membrane in which the target GPCR is embedded or may be present in another membrane printed in conjunction with the target membrane on a microspot. Or, a GPCR microarray may be pre-labeled by incorporating a label on an exposed substrate in a defect in the printed microspot.
-
Citations
25 Claims
-
1. A reference prelabeled G-protein coupled receptor membrane array comprising a plurality of assayable microspots, wherein one or more of the plurality of the microspots comprise:
-
a membrane comprising (i) a target G-protein coupled receptor embedded the membrane and (ii) a membrane component; and a labeled agent bound to the membrane component. - View Dependent Claims (2, 3)
-
-
4. A reference prelabeled G-protein coupled receptor membrane array comprising a plurality of assayable microspots, wherein one or more of the plurality of the microspots comprise:
-
a first membrane comprising a target G-protein coupled receptor embedded the membrane; a second membrane comprising a membrane component; a labeled agent bound to the membrane component; and wherein the first membrane is isolated from a cell overexpressing the G-protein coupled receptor and wherein the second membrane is isolated from a non-overexpressing cell. - View Dependent Claims (5, 6)
-
-
7. A method comprising:
-
Providing an isolated membrane from a cell overexpressing a G-protein coupled receptor, the membrane comprising a component; contacting the membrane with a labeled agent configured to bind the component to produce a pre-labeled membrane; and printing the pre-labeled membrane as a microspot on a substrate. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14)
-
-
15. A method comprising:
-
providing a first membrane from a cell overexpressing a G-protein coupled receptor; isolating a second membrane from a non-overexpressing cell, the second membrane comprising a component; contacting the second membrane with a labeled agent configured to bind the component to produce a pre-labeled second membrane; and printing the first and pre-labeled second membrane as a microspot on a substrate. - View Dependent Claims (16, 17, 18)
-
-
19. A method comprising:
-
contacting a microspot of a G-protein coupled receptor membrane array with labeled non-hydrolysable GTP analog; contacting the microspot with a first labeled agent configured to bind a membrane component other than the G-protein coupled receptor to produce a reference labeled membrane; contacting the microspot with a ligand suspected of being capable of functionally interacting with the G-protein coupled receptor; rinsing the microspot to remove unbound labeled non-hydrolysable GTP analog, unbound first labeled agent, and unbound ligand from the microspot; quantifying a first signal associated with the labeled non-hydrolysable GTP analog associated with the microspot; quantifying a second signal associated with the first labeled agent bound to the membrane component; and comparing the first signal with the second signal to determine the extent of the functional interaction of the agent with the G-protein coupled receptor. - View Dependent Claims (20, 21, 22)
-
-
23. A method comprising:
-
contacting a microspot of a G-protein coupled receptor membrane array with a first labeled agent; contacting the microspot with a second labeled agent configured to bind a membrane component; rinsing the microspot to remove unbound first labeled agent and unbound second labeled agent; quantifying a first signal associated with the first labeled agent associated with the microspot; quantifying a second signal associated with the second labeled agent bound to the membrane component; and comparing the first signal with the second signal to determine the extent of binding of the second labeled agent to membrane component. - View Dependent Claims (24, 25)
-
Specification