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Human Aβ1-40(Amyloid Beta 1-40) ELISA Kit

Citations(21)Uniprot : P05067

  • Cat.No.:E-EL-H0542

  • Reactivity: Human
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Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
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Product Details

Properties

Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Detection range 15.63-1000 pg/mL
Sensitivity 9.38 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Human Aβ1-40 in samples. No significant cross-reactivity or interference between Human Aβ1-40 and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Human Aβ1-40 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human Aβ1-40. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human Aβ1-40 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human Aβ1-40, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human Aβ1-40. You can calculate the concentration of Human Aβ1-40 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips 		-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL 		-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human Aβ1-40 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human Aβ1-40 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 50.60 152.10 454.70 45.80 156.80 436.80
Standard deviation 3.30 6.10 23.60 2.60 9.10 13.10
CV (%) 6.52 4.01 5.19 5.68 5.80 3.00

Recovery

The recovery of Human Aβ1-40 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 93-109 101
EDTA plasma (n=8) 85-96 91
Cell culture media (n=8) 92-108 100

Linearity

Samples were spiked with high concentrations of Human Aβ1-40 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database LinksSwissProt:   P05067
SynonymsAbeta 40, amyloid beta 40,Beta-amyloid protein 40,Aβ(1-40) ,Aβ40,Amyloid Beta 40
Research AreaCancer, Neuroscience

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
elisa assay procedure 3

3. Aspirate and wash the plate for 3 times
elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
elisa assay procedure 6

6. Add 50μL Stop Solution
elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. Cell Stem Cell (2020) IF: 20.86
    Interneuron Accumulation of Phosphorylated tau Impairs Adult Hippocampal Neurogenesis by Suppressing GABAergic Transmission

    DOI: 10.1016/j.stem.2019.12.015

    PMID: 31978364

    		 Sample: Cell culture supernatant
  2. Bioactive Materials (2021) IF: 14.593
    Modulating autophagic flux via ROS-responsive targeted micelles to restore neuronal proteostasis in Alzheimer's disease

    DOI: 10.1016/j.bioactmat.2021.09.017

    PMID: 34977433

    		 Sample: HT22 cell
  3. Autophagy (2019) IF: 11.059
    Activation of PPARA-mediated autophagy reduces Alzheimer disease-like pathology and cognitive decline in a murine model

    DOI: 10.1080/15548627.2019.1596488

    PMID: 30898012

    		 Sample: Tissue homogenate,Cell culture supernatant,Cell ly
  4. JOURNAL OF CONTROLLED RELEASE (2023) IF: 11.467
    Brain-neuron targeted nanoparticles for peptide synergy therapy at dual-target of Alzheimer's disease

    DOI: 10.1016/j.jconrel.2023.01.074
    Sample: hippocampal tissues
  5. AGING CELL (2021) IF: 9.304
    Platelet biomarkers identifying mild cognitive impairment in type 2 diabetes patients

    DOI: 10.1111/acel.13469

    PMID: 34528736

    		 Sample: plasma
  6. Cell Reports (2018) IF: 8.032
    CIP2A Causes Tau/APP Phosphorylation, Synaptopathy, and Memory Deficits in Alzheimer’s Disease

    DOI: 10.1016/j.celrep.2018.06.009
    Sample: Cell lysate
  7. Journal of Neuroinflammation (2022) IF: 8.3219
    Far infrared light irradiation enhances Aβ clearance via increased exocytotic microglial ATP and ameliorates cognitive deficit in Alzheimer’s disease-like mice

    DOI: 10.1186/s12974-022-02521-y
    Sample: brain
  8. TRAFFIC (2022) IF: 6.144
    AP2S1 regulates APP degradation through late endosome-lysosome fusion in cells and APP/PS1 mice

    DOI: 10.1111/tra.12874
  9. ANALYTICAL CHEMISTRY (2019) IF: 6.35
    Glycopeptide Nanofiber Platform for Aβ-Sialic Acid Interaction Analysis and Highly Sensitive Detection of Aβ

    DOI: 10.1021/acs.analchem.9b00377
    Sample: serum
  10. Frontiers in Cell and Developmental Biology (2021) IF: 6.684
    Periphery Biomarkers for Objective Diagnosis of Cognitive Decline in Type 2 Diabetes Patients

    DOI: 10.3389/fcell.2021.752753
    Sample: plasma
  11. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES (2022) IF: 6.953
    A monomeric polysaccharide from Polygonatum sibiricum improves cognitive functions in a model of Alzheimer's disease by reshaping the gut microbiota

    DOI: 10.1016/j.ijbiomac.2022.05.185

    PMID: 35661666

    		 Sample: hippocampus tissue
  12. JOURNAL OF CLINICAL PERIODONTOLOGY (2020) IF: 5.241
    Periodontitis and systemic markers of neurodegeneration: A case–control study

    DOI: 10.1111/jcpe.13267
    Sample: Serum
  13. Frontiers in Cellular Neuroscience (2022) IF: 5.505
    Alpha- and Gamma-Tocopherol Modulates the Amyloidogenic Pathway of Amyloid Precursor Protein in an in vitro Model of Alzheimer’s Disease: A Transcriptional Study

    DOI: 10.3389/fncel.2022.846459
    Sample: SH-SY5Y cell
  14. EXPERIMENTAL NEUROLOGY (2020) IF: 4.691
    Purified oleocanthal and ligstroside protect against mitochondrial dysfunction in models of early Alzheimer's disease and brain ageing

    DOI: 10.1016/j.expneurol.2020.113248

    PMID: 32084452

    		 Sample: Cell lysate
  15. Current Alzheimer Research (2019) IF: 3.271
    Genistein Decreases APP/tau Phosphorylation and Ameliorates Aβ Overproduction Through Inhibiting CIP2A

    DOI: 10.2174/1567205016666190830113420
    Sample: HEK-293T cells
  16. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2023) IF: 3.322
    The mitochondria-targeted small molecule SS31 delays progression of behavioral deficits by attenuating β-amyloid plaque formation and mitochondrial/synaptic deterioration in APP/PS1 mice

    DOI: 10.1016/j.bbrc.2023.02.076
    Sample: hippocampus
  17. JOURNAL OF PERIODONTOLOGY (2018) IF: 3.392
    Periodontitis is associated with systemic inflammation and vascular endothelial dysfunction in patients with lacunar infarct

    DOI: 10.1002/JPER.18-0560

    PMID: 30417380

    		 Sample: Serum
  18. JOURNAL OF TRACE ELEMENTS IN MEDICINE AND BIOLOGY (2021) IF: 3.849
    Influence of manganese exposure on cognitive function, plasma APP and Aβ levels in older men

    DOI: 10.1016/j.jtemb.2021.126788

    PMID: 34015662

    		 Sample: Plasma
  19. Scientific Reports (2020) IF: 3.998
    Comparative Effects of Alpha- and Gamma-Tocopherol on Mitochondrial Functions in Alzheimer’s Disease In Vitro Model

    DOI: 10.1038/s41598-020-65570-4
    Sample: Cell culture medium
  20. NEUROCHEMICAL RESEARCH (2019) IF: 2.782
    Resveratrol Abrogates Hypoxia-Induced Up-Regulation of Exosomal Amyloid-β Partially by Inhibiting CD147

    DOI: 10.1007/s11064-019-02742-3

    PMID: 30771155

    		 Sample: Tissue homogenate,Cell culture medium
  21. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY (2017) IF: 1.751
    Evaluation of the Expression of Amyloid Precursor Protein and the Ratio of Secreted Amyloid Beta 42 to Amyloid Beta 40 in SH-SY5Y Cells Stably Transfected with Wild-Type, Single-Mutant and Double-Mutant Forms of the APP Gene for the Study of Alzheimer’s D

    DOI: 10.1007/s12010-017-2468-6

    PMID: 28417423

    		 Sample: Cell culture supernatant
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Reviews/Q&A

  • Show all
  • Reviews
  • Q&A
Q

S***********nSubmitted [ Jan 29 2024 ]

Asked: Hi, I am conducting an ELISA using cell culture media (DMEM + 10% FBS), but the O.D value keeps coming out negative. What could be the problem?

Reply

A

adminSubmitted [ Jan 29 2024 ]

Answered: Hello Sungwoon, Thanks for your feedback. We will reply to you by email soon.

Q

D******************ASubmitted [ May 03 2023 ]

Asked: Hi, I am going to start ELISA using this kit for my extracted protein samples. I have 2 questions to ask. The first question is what is the storage condition of diluted wash buffer? Can I store the diluted wash buffer in room temperature or it should be stored at 4 degree Celsius for optimum function. My second question is, the manual states addition of Stop Solution follows the same order as the addition of Substrate Reagent. What does the order means?

Reply

A

adminSubmitted [ May 03 2023 ]

Answered: Dear Danesh, thanks for reaching out! I'll reply to you by email. Pls pay attention to it.

Q

D******************ASubmitted [ May 03 2022 ]

Asked: Hi, I am planning to buy this kit (E-EL-H0542) for amyloid-beta 1-40 detection from my fibroblast cell culture. The term cell-culture media used in this manual refers to the supernatant culture media, right? And to double confirm, we cannot use cell lysate for this kit, right? I hope you will help me with the proper sample method I can use along with this kit.

Reply

A

adminSubmitted [ May 03 2022 ]

Answered: Hi DANESH, Good morning~ "The term cell-culture media used in this manual refers to the supernatant culture media, right?" -yes, correct "And to double confirm, we cannot use cell lysate for this kit, right?" -this kit can detect Cell lysates as well as Cell culture supernatant. Any inquiry, pls feel free to contact me Thank you

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