Biology Reference
In-Depth Information
Gs/Gi pathway
Intracellular cAMP
level variation
LANCE cAMP (Perkin
Elmer)
Competition assay between endogenously
produced cAMP and acceptor-labeled cAMP for
binding on donor-labeled cAMP antibody
(TR-FRET signal decreases as endogenous
cAMP is produced)
van Borren
et al.
18
HTRF cAMP (Cisbio
Bioassays)
Competition assay between endogenously
produced cAMP and acceptor-labeled cAMP for
binding on donor-labeled cAMP antibody
(TR-FRET signal decreases as endogenous
cAMP is produced)
Thomsen
et al.
17
Comparison:
Gabriel
et al.
23
QRET-based cAMP assay
(Perkin Elmer)
Competition assay between labeled and
endogenously produced cAMP: soluble quenchers
reduce the signal of unbound europium-labeled
cAMP in solution, whereas the antibody-bound
fraction is fluorescent
Martikkala
et al.
24
ERK1/2
phosphorylation
Cellul'erk (Cisbio Bioassays) Sandwich immunoassay: detection of
phosphorylated ERK by the production of TR-
FRET signal between a donor-labeled anti-ERK
antibody and an acceptor-labeled anti-phospho-
ERK antibody
Thomsen
et al.
17
G protein-
dependent/
independent
pathways
Ras activation
Not commercially available
yet
Assay based on homogeneous quenching
resonance energy transfer technique (QRET):
terbium-labeled GTP binding to small GTPase
protein H-Ras protects the signal of the label from
quenching, whereas the signal of the nonbound
fraction of Tb-GTP is quenched by a soluble
quencher
Martikkala
et al.
25
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