Silencer® Select pre-designed siRNA oligonucleotides targeting IFITM3 were purchased from Ambion and tested: siRNA ID #s195033, #s195034, and #s195035 (sense sequence 5΄-CCCACGUACUCCAACUUCCtt −3΄ and antisense sequence 5΄-GGAAGUUGGAGUACGUGGGat-3΄). IFITM3 siRNA (#s195035) and scrambled siRNA (negative control) were used. siRNAs were transfected to HEK293-APP695 cells in triplicates as described previously50 . The conditioned medium was measured for secreted Aβ40 and 42 levels by using Aβ peptide multiplex kits (6E10, MSD) according to manufacturer’s instructions. To check cell viability, Alamar Blue (AbD Serotec) was added to the cells and incubated50 . Transferred medium in a 96-well black polystyrene microplate (Costar) was read for fluorescence (530–560 nm excitation/ 590 nm emission) by EnVision Plate Reader (PerkinElmer). Protein knock-down levels were measured in cell lysates by WB. siRNA transfection in HEK293-NotchΔE cells was performed as described above. Levels of protein knock-down, NotchΔE and NICD were measured in cell lysates by WB.
Aβ peptide multiplex kits 6e10
Manufactured by MSD
The Aβ peptide multiplex kits (6E10) are research tools used to detect and quantify Aβ peptides. These kits utilize antibody-based detection methods, such as 6E10, to measure the levels of different Aβ species in biological samples. The core function of these kits is to provide researchers with a standardized and reliable way to analyze Aβ peptide levels, which is important for various applications in neurodegenerative disease research.
Automatically generated - may contain errors
2 protocols using aβ peptide multiplex kits 6e10
1
Silencing IFITM3 Impacts Amyloid-beta Secretion
2
Silencing IFITM3 Impacts Amyloid-beta Secretion
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Silencer® Select pre-designed siRNA oligonucleotides targeting IFITM3 were purchased from Ambion and tested: siRNA ID #s195033, #s195034, and #s195035 (sense sequence 5΄-CCCACGUACUCCAACUUCCtt −3΄ and antisense sequence 5΄-GGAAGUUGGAGUACGUGGGat-3΄). IFITM3 siRNA (#s195035) and scrambled siRNA (negative control) were used. siRNAs were transfected to HEK293-APP695 cells in triplicates as described previously50 . The conditioned medium was measured for secreted Aβ40 and 42 levels by using Aβ peptide multiplex kits (6E10, MSD) according to manufacturer’s instructions. To check cell viability, Alamar Blue (AbD Serotec) was added to the cells and incubated50 . Transferred medium in a 96-well black polystyrene microplate (Costar) was read for fluorescence (530–560 nm excitation/ 590 nm emission) by EnVision Plate Reader (PerkinElmer). Protein knock-down levels were measured in cell lysates by WB. siRNA transfection in HEK293-NotchΔE cells was performed as described above. Levels of protein knock-down, NotchΔE and NICD were measured in cell lysates by WB.
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