Biological descriptionγ-Secretase inhibitor. Inhibits Notch signalling in vitro. Inhibits total Aβ and Aβ42 production in human primary cultures (IC50 values are 115 and 200 nM, respectively) with no effect on APPα and APPβ levels. Blood-brain barrier permeable.
Chemical nameN-[N-(3,5-Difluorophenacetyl-L-alanyl)]-(S)-phenylglycine t-butyl ester
Storage instructionsStore at +4°C. The product can be stored for up to 12 months.
Solubility overviewSoluble in DMSO to 100 mM
Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Functional Studies - DAPT (ab120633)
ab81283 staining AKT1 (phospho S473) in MCF7 cells treated with DAPT (ab120633), by ICC/IF. Decrease in expression of AKT1 (phospho S473) correlates with increased concentration of DAPT, as described in literature. The cells were incubated at 37°C for 24h in media containing different concentrations of ab120633 (DAPT) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81283 (1/200 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Functional Studies - DAPT (ab120633)This image is courtesy of an Abreview submitted by a verified customer
Kc167 cells were treated with varying concentrations of DAPT (ab120633) in DMSO for 16hr at 25°C; DMSO only was used as the negative control. The cells were further incubated for 30 minutes with 4mM EGTA in PBS (in the presence of DAPT), and were then lysed for analysis. To measure Notch activity, Notch targets E(spl)mß-HLH and E(spl)m3-HLH mRNA levels were assayed. Data shows the fold change of mRNA levels of E(spl)mß-HLH and E(spl)m3-HLH under different conditions, normalised to DMSO treatment (negative control). Notch activation by EGTA is abrogated by treatment with DAPT.
Kanungo J et al. The Notch signaling inhibitor DAPT down-regulates cdk5 activity and modulates the distribution of neuronal cytoskeletal proteins. J Neurochem106:2236-48 (2008).
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