Patent ID: 11867698
Assignee: LIFE TECHNOLOGIES CORPORATION
Field: Measurement (Instruments)
Classification: CPC G  C  Y | IPC C  G

Claim 12:
13. A method for detecting receptor-mediated endocytosis, the method comprising:
(a) conjugating a target molecule to a fluorescent pH-sensitive dye of Formula I:, wherein,
R1, R2, R3 and R6 are each independently H, Z, or an electron donating group (EDG), provided that R1 and R2 are not hydroxyl or thiol or their deprotonated forms;
R4 is selected from the group consisting of H, alkyl, substituted alkyl, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, a reactive group, a carrier molecule, heterocyclyl, and substituted heterocyclyl;
R5 is independently selected from the group consisting of Y, alkyl, substituted alkyl, alkenyl, substituted alkenyl, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, a reactive group, a carrier molecule, heterocyclyl, and substituted heterocyclyl;
X is a fluorophore;
Y is ═CRbRc or ═CRbRd;
Z is —ORc, —SRc, —NRbRc;
Rb is H, alkyl, or substituted alkyl;
Rc is alkyl or substituted alkyl; and
Rd is amino or substituted amino;
or a stereoisomer, tautomer, or salt thereof to form a target conjugate;
(b) contacting the target conjugate with a cell to form a contacted cell;
(c) incubating the contacted cell to form an incubated solution;
(d) illuminating the incubated solution to form an illuminated solution; and
(e) detecting fluorescent emissions from the illuminated solution;
wherein fluorescent emissions indicate receptor-mediated endocytosis of the target molecule and wherein no quenching step is performed before the detecting step.