#Gamma control ic serial#
#Gamma control ic plus#

However, in the presence of substrate, ELN475516 displaces 50% of the active site probe at a concentration equivalent to approximately 67-fold its Gamma APP IC 50. (d) ELN-475516 does not displace the active site probe at concentrations ranging up to 2,000X its IC 50 in the gammaAPP assay. In the presence of substrate, ELN318463 displaces approximately 50% of the active site probe at a concentration of approximately 2,000-fold its IC 50 in the Gamma APP assay. (c) No displacement of active site probe is observed by ELN-318463 in the absence of substrate. In the presence of added APP, the sulfonamides displace the active site probe in a dose-dependent manner. Substrate was added to enzyme concurrently with test compound and affinity ligand. Hence, the ability of sulfonamides to displace the active site directed probe was tested in the absence (c and d, left side ) or presence (c and d, right side) of 1 Km substrate (MBP-C125sw). Previous observations revealed substrate concentration affects the potency of sulfonamides in cell and enzyme assays (unpublished). At higher concentrations, a modest dose-dependent effect of LY411575 to displace the active-site-directed compound is observed on both Western blots.

There is no significant displacement of the active site directed compound by at concentrations below 200-fold its IC 50 in the enzyme assay. (b) LY411575 was tested for its ability to displace active site isostere at concentrations ranging up to 1000-fold its IC 50 in the Gamma APP enzyme assay. In subsequent experiments, a 200-fold excess of L-685,458 was employed as a positive control. Saving the file at this location will make it appear in the list of profiles the next time you open System Preferences and go to the Screens panel. By default, it’ll save the file at the standard location for profiles in your user account. (a) The biotinylated isostere is displaced by its non-biotinylated analog L-685,458 in a concentration dependent manner. Choose Export Color Profile from the Gamma menu.

#Gamma control ic serial#

Serial dilutions of affinity captured enzyme were included on the gels to provide a standard curve for densitometric quantitation of test samples following autoradiography of the western blots. The binding assay was carried out using 50 nM affinity probe together with different concentrations of competing compounds. The eluted enzyme captured on neutravidin beads was detected on western blots with antibodies recognizing either PS-1 amino-terminal fragment (PS1-NTF), or Nicastrin (Nct).

#Gamma control ic plus#

A competitive binding assay employing a biotin conjugated active site isostere to capture gamma secretase from a partially purified cell extract, plus or minus co-incubation with increasing concentrations of reference or test gamma secretase inhibitors, was developed to characterize inhibitor binding site. Binding site analysis by competitive displacement of biotinylated active site isostere.