Amyloid beta1-40 Kit
Amyloid beta1-40 quantification from cells and supernatants
Amyloid ß1-40 and ß1-42 peptides are involved in the appearance of Alzheimer's Disease symptoms. The production of these two amyloid peptides is regulated by the activity of 2 enzymes, ß and γ secretase, which cleave the amyloid precursor protein. This kit is intended to measure amyloid ß1-42 peptides in cell supernatant or in whole cells. The assay is a fast alternative to ELISA, thanks to the easy implementation of our Add and Read method.
The human amyloid beta 1-42 assay measures human amyloid 1-42 peptide in a cell or tissue supernatant or lysate. The assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the human beta amyloid 1-42 peptide. In presence of human beta 1-42 peptide in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing any changes caused by experimental variability under a no-wash assay format.
Final assay volume
Time to results
Overnight at RT
40 to 2,000 pg/mL
Cell supernatant, Cell Lysate (LB#1)
|Sample||Mean [amyloid 1-42] (pg/mL)||CV|
Each of the 3 samples was measured 24 times, and the %CV was calculated for each sample.
|Sample||Mean [amyloid 1-42] (pg/mL)||CV|
Each of the samples was measured in 4 independent experiments by 4 different operators, and the % CV was calculated for each sample.
[Standard amyloid 1-42] (pg/mL)
[native amyloid 1-42] added (pg/mL)
[amyloid 1-42] expected (pg/mL)
[amyloid 1-42] measured (pg/mL)
Two levels of recombinant protein (400 and 850 pg/mL) were added to 3 dilutions of native sample from HEK293 APPsw, cell supernatants and the expected concentrations were compared to those measured in order to compute antigen recoveries.
C expected (pg/mL)
C obtained (pg/mL)
Dilution experiments were performed in regular microtubes. Low binding microtubes did not shown any significant changes in dilutional linearity (data not shown). Samples were from C.elegans cell extracts diluted in the kit diluent DIL5. The recovery % obtained from these experiments show good dilutional linearity of the assay.
Amyloid beta 1-42
Amyloid beta 3-42
Amyloid beta 1-38
Amyloid beta 1-43
Amyloid beta 17-42
Amyloid beta 1-40
Cross reactivities were determined by using standard curves of recombinant peptides in the kit Diluent, with concentrations ranging from 60 to 2000 pg/mL. interference was tested by spiking 6 increasing concentrations of peptides (from 125 up to 4000 pg/mL) in a sample containing an amyloid beta 1-42 peptide concentration of 350 pg/mL.
 Oules , B., D. Del Prete , B. Greco, X. Zhang, I. Lauritzen, J. Sevalle , S. Moreno, P. Paterlini-Brechot , M. Trebak , F. Checler , F. Benfenati & M. Chami (2012) Ryanodine receptor blockade reduces amyloid-beta load and memory impairments in Tg2576 mouse model of Alzheimer Disease. J Neurosci , 32, 11820-34.
Amyloid ß1-40 and ß1-42 peptides are involved in the appearance of Alzheimer's disease symptoms. The production of these two amyloid peptides is regulated by the activity of 2 enzymes, ß and γ secretase, which cleave the amyloid precursor protein. In normal conditions, the Amyloid Precursor Protein (APP) is processed through the non –amyloidogenic pathway, the alpha-secretase cleavage generating soluble APP involved in normal neuronal function. However, in pathological conditions, there is an abnormal cleavage of APP by the beta-secretase followed by the gamma-secretase which release the amyloid peptides that are prone to aggregation. This amyloidogenic pathway can result in plaque formation, impairs neuronal function and can lead to neurodegeneration. Measurement of secreted beta amyloid peptides is of critical importance in the evaluation of the amyloidogenic pathway activity in Alzheimer Disease cellular models.
CTFa/b: Carboxy-Terminal Fragment alpha/beta - APP: Amyloid Precursor Protein - ICD: APP Intracellular Domain - P3: p3 protein = amyloid beta 17-42 (non amyloidogenic)
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