PMID- 9025951
OWN - NLM
STAT- MEDLINE
DCOM- 19970407
LR  - 20131121
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 244
IP  - 2
DP  - 1997 Jan 15
TI  - A chemiluminescent microtiter plate assay for sensitive detection of protein 
      kinase activity.
PG  - 340-6
AB  - A chemiluminescent protein kinase assay using biotinylated substrate peptides 
      captured on a streptavidin-coated microtiter plate and monoclonal antibodies to 
      detect their phosphorylation is described. Assay conditions were optimized and 
      validated for sensitive measurement of protein kinase A, protein kinase C, 
      Ca2+/calmodulin-dependent protein kinase II (CAM-KII), receptor interacting 
      protein, and src activities. The newly developed chemiluminescent assay has 
      several advantages over currently used radioactive or colorimetric methods. It is 
      highly sensitive at low enzyme and substrate concentrations and high, close to 
      physiological ATP levels. It is fast, simple to perform and amenable to 
      automation and high-throughput drug screening. The assay is also robust, 
      exhibiting minimum interference from solvents and test substances from various 
      sources. Overall, among the presently available methods for the detection of 
      protein kinase activity, chemiluminescence was found to provide the highest 
      sensitivity under conditions most closely mimicking the intracellular 
      environment. This assay is expected to be useful in both academic and industrial 
      laboratories, especially in identifying novel classes of protein kinase 
      inhibitors.
FAU - Lehel, C
AU  - Lehel C
AD  - Tularik Inc., South San Francisco, California 94080, USA.
FAU - Daniel-Issakani, S
AU  - Daniel-Issakani S
FAU - Brasseur, M
AU  - Brasseur M
FAU - Strulovici, B
AU  - Strulovici B
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Glial Fibrillary Acidic Protein)
RN  - 0 (Proteins)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.10.2 (src-Family Kinases)
RN  - EC 2.7.11.1 (Receptor-Interacting Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.1 (Ripk1 protein, mouse)
RN  - EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
SB  - IM
MH  - Adenosine Triphosphate/metabolism
MH  - Animals
MH  - Apoptosis
MH  - Colorimetry
MH  - Cyclic AMP-Dependent Protein Kinases/*analysis
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Glial Fibrillary Acidic Protein/analysis
MH  - *Luminescent Measurements
MH  - Mice
MH  - Phosphorylation
MH  - Proteins/analysis
MH  - Receptor-Interacting Protein Serine-Threonine Kinases
MH  - src-Family Kinases/analysis
EDAT- 1997/01/15 00:00
MHDA- 1997/01/15 00:01
CRDT- 1997/01/15 00:00
PHST- 1997/01/15 00:00 [pubmed]
PHST- 1997/01/15 00:01 [medline]
PHST- 1997/01/15 00:00 [entrez]
AID - S0003-2697(96)99894-2 [pii]
AID - 10.1006/abio.1996.9894 [doi]
PST - ppublish
SO  - Anal Biochem. 1997 Jan 15;244(2):340-6. doi: 10.1006/abio.1996.9894.