pcDNA3-flag-hAPP695 was as previously described [50 (link)]. pcDNA4/V5-His-hAICD59 expressing the C-terminal 59 amino acid was subcloned from full-length hAPP695. SH-SY5Y cells were obtained from ATCC and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% (v/v) FBS. SH-SY5Y cells were transfected with various plasmids using the Lipofectamine 2000 reagent (Invitrogen) according to the instructions of the manufacturer. Two days after transfection, the cells were used for experiments. The SH-SY5Y cell line-overexpressing APP (SH-SY5Y-APP) was generated by transfection with a pcDNA4 plasmid construct (Invitrogen) carrying APP695 and a Zeocin resistance gene followed by selection for Zeocin resistance. To inhibit AICD degradation, the cells were treated with 10 mM NH4Cl overnight in the culture medium [57 (link)]. L-685,458 (5 μM final concentration; Tocris, catalog no. 2627) or BMS 299897 (1 μM final concentration; Tocris, catalog no. 2870) were administered to the cells in culture medium for 24 h.
Bms 299897
Manufactured by Bio-Techne
The BMS 299897 is a lab equipment product offered by Bio-Techne. It is designed for laboratory use, but a detailed description of its core function is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
L 685 458, by Bio-Techne (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 546, by Thermo Fisher Scientific (1 mentions)
Gi254023x, by Merck Group (1 mentions)
Tapi 1, by Merck Group (1 mentions)
Mouse anti gfp antibody, by Abcam (1 mentions)
Batimastat, by Merck Group (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Azd3839, by Selleck Chemicals (1 mentions)
2 protocols using bms 299897
1
Characterizing APP and AICD Regulation
2
Antibody and Secretase Inhibitor Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mouse anti-GFP antibody (1:2000) was purchased from Abcam; Rabbit anti-FOXO3a (75D8) antibody (1:300) and rabbit anti-pAkt S473 antibody (1:500) were obtained from Cell Signaling Technology; Rabbit anti-Amyloid Precursor Protein, C-Terminal amino acid 676-695 (1:2000; A8717) was purchased from Sigma. Goat antirabbit antibody Alexa Fluor 546 (1:600) and goat anti-mouse antibody Alexa Fluor 488 (1:2000) were purchased from Invitrogen. 4',6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI; 1mg/ml) from ThermoFisher Scientific. TAPI-1, Batimastat and β-secretase inhibitor IV (β-IV) were purchased from Merck Millipore, DAPT and BMS299897 were from Tocris Bioscience, AZD3839 from Selleckchem and GI 254023X from Sigma Aldrich. All secretase inhibitors were reconstituted in dimethylsulphoxide (DMSO) at stock solutions of 1-10 mM as recommended by the manufacturer.
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