NewEastBio Monoclonal cGMP ELISA Kit 实验分析级cAMP ELISA试剂盒

96 Wel l Ki t

Product Description
Guanosine 3', 5'-cyclic monophosphate (cyclic GMP; cGMP) playscritical
regulatory roles in many physiological processes. cGMP is producedfrom
GTP by guanylyl cyclases and is degraded byphosphodiesterases.
Stimulation of guanylyl cyclases or inhibition ofphosphodiesterases can
increase cellular cGMP concentrations. Inhibitors of thecGMP-specific
phosphodiesterases are used for
treating human diseases. For
example, inhibitors of cGMP specific
phosphodiesterase type 5 (such as
Viagra and Cialis) are used for
treating erectile dysfunction.
To screen for inhibitors of
phosphodiesterases or stimulators of
guanylyl cyclases, it is essential to
have a fast, sensitive, selective and
reproducible method to measure the
cGMP concentrations. This is
especially true for the initial
screenings given the possible weaker
effects of larger pools of compounds.
Currently available other ELISA kits measuring cGMP levels arebased on
the non-affinity-purified polyclonal anti-cGMP antibody. Despitethe
claimed selectivity, these polyclonal anti-cGMP antibodies displaycertain
cross-reactivity with cAMP or GTP. In most cell types, cGMP ispresent at
levels ~100 fold lower than cAMP.
NewEast Biosciences cGMP ELISA kit is based on the uniquemouse
monoclonal anti-cGMP antibody. This monoclonal anti-cGMPantibody
displays >108 fold of selectivity over cAMP, GTP, and othernucleoside
analogues. NewEast Biosciences cGMP ELISA kit providessignificantly
improved sensitivity and selectivity over other kits based onpolyclonal
anti-cGMP antibodies. Our monoclonal anti-cGMP antibody-based
ELISA kit also avoids the batch-to-batch variations associatedwith
polyclonal antibody productions from animals, thus providingthe
reproducibility in the long run.
Furthermore, while polyclonal anti-cGMP antibodies used in otherELISA
kits have higher affinity for acetylated cGMP than non-acetylatedcGMP,
NewEast Biosciences monoclonal anti-cGMP antibody has similar
affinities to non-acetylated and acetylated cGMP molecules.Therefore,
acetylation treatments of samples and standards are not needed inNewEast
Biosciences cGMP ELISA kit. This significantly reduces the time forthe
assay. The avoidance of organic reagents used in the acetylationprocess
provides a safe and healthy work environment.
Principle Outline
NewEast Biosciences cGMP ELISA Kit is a competitive immunoassayto
measure the cGMP levels, either from cell extracts or from invitro
guanylyl cyclase assays. Briefly, multi-well plates are coatedwith
goat-anti-mouse serum. cGMP in cell extracts or in in vitroguanylyl
cyclase assays will competitively bind to the monoclonalanti-cGMP
antibody in the presence of fixed amounts of cGMP-conjugated
horse-radish peroxidase. Known amounts of cGMP are used asstandards
to generate the calculation curve. After a short incubation, theexcess
reagents are washed away and substrate is added. The multiwellplates are
then read on a microplate reader at 450 nm. The intensity of theyellow
color is inversely proportional to the concentration of cGMP insamples.
The measured optical density is used to calculate the concentrationof
cGMP in samples based on the curve from the cGMPstandards.