This report is the first of its kind on screening of Indian medicinal
plants in ROCK-II inhibition assay using HTRFs KinEASETM STK S2 Kit and correlating the data with traditional literature which reports
the utility of these plants in Vajikarana chikitsa. ROCK-II inhibition
assay was performed by HTRF technique, a homogeneous assay
procedure which was preferred over ELISA based technique since it
does not involve several washing steps as required in assay based on
ELISA method. Besides, herbal extracts having antioxidant effect
interfere with horseradish peroxidase-based detection system in
Enzyme-linked immunosorbent assay (ELISA) and can lead to false
positive results (Dufau et al., 2008). The ROCK-II inhibition assay
was standardized for the optimum concentration of the individual
components of the kit, viz., substrate, ATP, SA-XL665 and validated
with Y-27632, a standard inhibitor, prior to screening of the plant
extracts (data not given). Varying ROCK-II inhibition was exhibited
by 15 different extracts, wherein the corresponding vehicle control
(2.5% DMSO) did not show any enzyme inhibition.
This report is the first of its kind on screening of Indian medicinalplants in ROCK-II inhibition assay using HTRFs KinEASETM STK S2 Kit and correlating the data with traditional literature which reportsthe utility of these plants in Vajikarana chikitsa. ROCK-II inhibitionassay was performed by HTRF technique, a homogeneous assayprocedure which was preferred over ELISA based technique since itdoes not involve several washing steps as required in assay based onELISA method. Besides, herbal extracts having antioxidant effectinterfere with horseradish peroxidase-based detection system inEnzyme-linked immunosorbent assay (ELISA) and can lead to falsepositive results (Dufau et al., 2008). The ROCK-II inhibition assaywas standardized for the optimum concentration of the individualcomponents of the kit, viz., substrate, ATP, SA-XL665 and validatedwith Y-27632, a standard inhibitor, prior to screening of the plantextracts (data not given). Varying ROCK-II inhibition was exhibitedby 15 different extracts, wherein the corresponding vehicle control(2.5% DMSO) did not show any enzyme inhibition.
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