Highlights |
Fully pre-configured sandwich ELISA format providing convenience, high sensitivity and selectivity. Strip plate configuration allowing use and storage of separated sections of the ELISA plate. Affinity purified polyclonal capture antibody on plate. Distinct HRP-conjugated. detection antibody. Highly sensitive detection of sAPPalpha in cell culture medium and biological fluids. Detection ranges are 1-180 pg/ml depending on species. Detects the soluble alpha-secretase cleavage product of amyloid precursor protein in cell culture medium and biological fluids, designated sAPPalpha, after centrifugation and sedimentation of cells. Assays are done with the supernatant fraction to quantitate sAPPalpha. This ELISA kit can also detect full length APP (amyloid precursor protein) in cell and tissue lysates after dissolution from the membrane. To distinguish sAPPalpha from membrane bound APP, first sediment cells and collect the supernatant to assay sAPPalpha. Then use a non-denaturing cell lysis buffer to release and solubilize full length APP from the cell pellet to assay the remaining uncleaved APP. Rapid assay with colorimetric detection (measure reaction at 450nm). Total incubation with ELISA plate and HRP antibody is 1 hr at 37C or 2 hr at RT. |
Background |
sAPPalpha is the extracellular soluble fragment of amyloid precursor protein cleaved by "alpha-secretase." Generation of sAPPalpha by "alpha-secretase" prevents development of fibrillogenic amyloid-beta since the cleavage site is within the amyloid-beta molecule. The generation of sAPPalpha may therefore be neuroprotective. sAPPalpha may function in neuronal excitability, synaptic plasticity, neurite outgrowth, synaptogenesis, and cell survival. sAPPalpha also appears to modulate potassium channels, N-methyl-D-aspartate receptors, and the transcription factor NF kappa B1. |
References |
1. Lopez-Perez E, Dumanchin C, Czech C, Campion D, Goud B, et al. 2000. Overexpression of Rab11 or constitutively active Rab11 does not affect sAPPalpha and Abeta secretions by wild-type and Swedish mutated betaAPP-expressing HEK293 cells. Biochem Biophys Res Commun 275: 910-5
2. Maillet M, Robert SJ, Cacquevel M, Gastineau M, Vivien D, et al. 2003. Crosstalk between Rap1 and Rac regulates secretion of sAPPalpha. Nat Cell Biol 5: 633-9
3. Mousavi M, Hellstrom-Lindahl E. 2009. Nicotinic receptor agonists and antagonists increase sAPPalpha secretion and decrease Abeta levels in vitro. Neurochem Int 54: 237-44
4. Siemes C, Quast T, Klein E, Bieber T, Hooper NM, Herzog V. 2004. Normalized proliferation of normal and psoriatic keratinocytes by suppression of sAPPalpha-release. J Invest Dermatol 123: 556-63
5. Smirnov A, Trupp A, Henkel AW, Bloch E, Reulbach U, et al. 2009. Differential processing and secretion of Abeta peptides and sAPPalpha in human platelets is regulated by thrombin and prostaglandine 2. Neurobiol Aging 30: 1552-62
6. Talamagas AA, Efthimiopoulos S, Tsilibary EC, Figueiredo-Pereira ME, Tzinia AK. 2007.
Abeta(1-40)-induced secretion of matrix metalloproteinase-9 results in sAPPalpha release by association with cell surface APP. Neurobiol Dis 28: 304-15 |