High Molecular Amyloid β Oligomer ELISA Kit Wako Ver.2
- for Immunochemistry
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
- Label
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96Tests
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Document
Kit component
96 tests
Antibody (BAN50)-coated Plate | 1 plate |
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Standard Solution (10 nM) | 100 μL |
Buffer | 60 mL |
Biotin-conjugated Antibody(BAN50) Solution | 100 μL |
Peroxidase-conjugated Streptavidin Solution | 100 μL |
Luminescent Reagent 1 | 6 mL |
Luminescent Reagent 2 | 6 mL |
Wash Solution (10 x) | 100 mL |
Sample Buffer for Serum/Plasma | 50 mL |
Plate Seal | 4 sheets |
Overview
Features
- Detects in serum and plasma, CSF sample
- Specifically reactive with ≧9mer Aβ oligomer
Performance
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Standard curve range 0.41-100 pM (Cerebrospinal fluid)
0.16-40 pM (serum or plasma(EDTA))Reactive Aβ oligomers ≧9mer Aβ oligomer Sample Serum, Plasma(EDTA),
Cerebrospinal fluid(CSF),
In vitro Aβ oligomerCross-reactivity Human Sample volume CSF:25μL(4 times dilution)
Serum, plasma:50μL(2 times dilution)Assay time 4.5 hours Detection method Luminescence method** - Calculated based on 16-mer MAP peptide.
- * Requires luminescence plate reader for measurement.
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Assay principal
This ELISA kit is designed to measure ≧ 9-mer Aβ oligomer specifically by using same Aβ antibody (BAN50)
for capture and detection3)4). As a standard, a 16-mer MAP peptide with 16 epitopes of anti Aβ antibody (BAN50) is used4).
Specificity for Aβ oligomers in the ELISA
Aβ monomers were incubated the solution at 4℃ overnight in order to synthesize various size of Aβ oligomers. They were separated Fraction A and Fraction B by ultrafiltration. The fractions were analyzed by western blotting and the Aβ oligomer ELISA.
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Western blotting using Aβ antibody(BAN50)
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Aβ oligomer ELISA
A strong signal was exhibited in the fraction A containing 9-mer and more (>30 kDa), whereas few signal was detected in fraction B containing 8-mer and smaller (<30 kDa).
Example of measurements
Example of measurement 1 - spiked recovery test and dilution linearity -
Human plasma | Not spiked | +0.5pM Std. | +0.1pM Std. | +5pM Std. | +30pM Std. | ||||
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Assay value | Assay value | Recovery | Assay value | Recovery | Assay value | Recovery | Assay value | Recovery | |
1.85pM | 2.36pM | 102% | 2.85pM | 100% | 7.11pM | 105% | 31.2pM | 97.8% | |
Human serum | Not spiked | +0.5pM Std. | +0.1pM Std. | +5pM Std. | +30pM Std. | ||||
Assay value | Assay value | Recovery | Assay value | Recovery | Assay value | Recovery | Assay value | Recovery | |
1.58pM | 2.07pM | 98% | 2.59pM | 101% | 6.64pM | 101% | 31.9pM | 101% |
Human CSF1 | Not spiked | +3pM Std. | +6pM Std. | +15pM Std. | |||
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Assay value | Assay value | Recovery | Assay value | Recovery | Assay value | Recovery | |
5.16pM | 8.01pM | 95% | 10.6pM | 91% | 18.1pM | 86% | |
Human CSF2 | Not spiked | +3pM Std. | +6pM Std. | +15pM Std. | |||
Assay value | Assay value | Recovery | Assay value | Recovery | Assay value | Recovery | |
29.1pM | 38.8pM | 97% | 44.0pM | 99.3% | 59.9pM | 88% |
Example of measurement 2 - CSF sample -
Cerebrospinal fluids(CSF) of patients with Alzheimer disease (AD) and Mild Cognitive Impairment (MCI), non-dementia patients(control), were measured with our ELISA kit and competitor’s kit.
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Age Sex MMSE※3 CSF
Aβ oligomer(pM)Control
n=569 M 30 2.37 60 F 30 1.65 59 M 30 2.82 46 M 28 2.24 60 F 29 1.34 MCI
n=569 F 24 3.6 76 M 23 2.62 82 M 26 2.33 69 F 23 2.67 73 F 25 2.34 AD
n=879 F 10 3.56 88 M 24 3.13 62 M 16 4.13 78 M 13 3.95 63 M 14 3.88 74 F 16 3.25 76 M 19 3.69 84 M 16 4.34 - ***MMSE score:diagnosis Inquiry test score of cognitive function
30-28 Normal
27-24 Suspected of mild cognitive impairment
≦23 Suspected of dementia
- ***MMSE score:diagnosis Inquiry test score of cognitive function
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- There was a significant difference between Control, MCI and AD.
- High molecular Aβ oligomer correlated with MMSE score.
Alias
- Aβoligomer ELISA Kit
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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