Multiplex Assay Kit for Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array)

AAA; ABeta; ABPP; AD1; APPI; CVAP; PN2; CTFgamma; Peptidase Nexin-II Alzheimer Disease; Peptidase Nexin-II; Amyloid Beta(A4)Precursor Protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 96
EDTA plasma(n=5) 86-105 90
heparin plasma(n=5) 90-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Amyloid Precursor Protein (APP) ,etc. by CBA (Cytometric Bead Array) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 78-105% 90-98% 93-101% 84-104%
EDTA plasma(n=5) 87-104% 78-91% 90-104% 83-96%
heparin plasma(n=5) 88-102% 78-104% 78-101% 81-97%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Neurochemistry International Bilobalide regulates soluble amyloid precursor protein release via phosphatidyl inositol 3 kinase-dependent pathway ScienceDirect: S0197018611001641
Neuroscience Substitution of membrane cholesterol with β-sitosterol promotes nonamyloidogenic cleavage of endogenous amyloid precursor protein Pubmed: 23707801
OncoImmunology PROOF COVER SHEET
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Neurotoxicology Involvement of hippocampal agmatine in β1-42 amyloid induced memory impairment, neuroinflammation and BDNF signaling disruption in mice Pubmed: 32522471
Metab Brain Dis Probiotics ameliorates LPS induced neuroinflammation injury on Aβ 1–42, APP, γ-β secretase and BDNF levels in maternal gut microbiota and fetal … Pubmed:35312928
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