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ACH(Acetylcholine) ELISA Kit

  • Cat.No.:E-EL-0081

  • Reactivity: Universal

To Purchase E-EL-0081

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
Qty:

Product Details

Properties

Assay type Competitive-ELISA
Format 96T/48T
Assay time 2.0h
Detection range 15.63-1000 pg/mL
Sensitivity 9.38 pg/mL
Sample type &Sample volume serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes ACH in samples. No significant cross-reactivity or interference between ACH 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 ACH concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ACH. During the reaction, ACH in the sample or standard competes with a fixed amount of ACH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to ACH. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of ACH in tested samples can be calculated 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℃
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 ACH were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level ACH 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) 50.90 124.30 438.10 55.30 116.50 429.80
Standard deviation 2.60 6.10 13.60 3.50 5.70 15.50
CV (%) 5.11 4.91 3.10 6.33 4.89 3.61

Recovery

The recovery of ACH 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) 89-102 96
EDTA plasma(n=8) 88-102 93
Cell culture media(n=8) 87-102 94

Linearity

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

Target Information

Assay Procedures

elisa assay procedure 1

1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

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

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

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

Citations

  1. Advanced Science (2021) IF: 16.806
    Neuronal Induction of Bone-Fat Imbalance through Osteocyte Neuropeptide Y

    DOI: 10.1002/advs.202100808

    PMID: 34719888

    Sample: marrow-depleted bone
  2. Food & Function (2022) IF: 5.396
    A randomised, double-blind, placebo-controlled trial of Bifidobacterium bifidum CCFM16 for manipulation of gut microbiota and relief from chronic constipation

    DOI: 10.1039/D1FO03896F

    Sample: serum
  3. Frontiers in Psychiatry (2022) IF: 5.435
    Efficacy of a novel proprietary dietary supplement (TRI 360TM) on psychological symptoms and stress-related quality of life in adult subjects: A randomized controlled clinical trial

    DOI: 10.3389/fpsyt.2022.919284

    PMID: 36032243

    Sample: serum
  4. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2021) IF: 5.924
    Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC

    DOI: 10.3390/ijms22137086

    PMID: 34209365

    Sample: WJ-MSC cells
  5. SAUDI JOURNAL OF BIOLOGICAL SCIENCES (2021) IF: 4.219
    Neuroprotective efficacy of 4-Hydroxyisoleucine in experimentally induced intracerebral hemorrhage

    DOI: 10.1016/j.sjbs.2021.07.010

    PMID: 34764759

    Sample: brain
  6. MOLECULES (2022) IF: 4.412
    Guggulsterone Mediated JAK/STAT and PPAR-Gamma Modulation Prevents Neurobehavioral and Neurochemical Abnormalities in Propionic Acid-Induced Experimental Model of Autism

    DOI: 10.3390/molecules27030889

    Sample: brain
  7. CHEMOSPHERE (2018) IF: 4.427
    Study on neurotoxicity of dinotefuran, thiamethoxam and imidacloprid against Chinese lizards (Eremias argus)

    DOI: 10.1016/j.chemosphere.2018.11.016

    PMID: 30419375

    Sample: Plasma
  8. Life-Basel (2022) IF: 3.251
    Survival and Neurogenesis-Promoting Effects of the Co-Overexpression of BCLXL and BDNF Genes on Wharton’s Jelly-Derived Mesenchymal Stem Cells

    DOI: 10.3390/life12091406

    Sample: WJ-MSC cell
  9. International Journal of Environmental Research and Public Health (2021) IF: 3.39
    Polystyrene Microplastics Exposure: An Insight into Multiple Organ Histological Alterations, Oxidative Stress and Neurotoxicity in Javanese Medaka Fish (Oryzias javanicus Bleeker, 1854)

    DOI: 10.3390/ijerph18189449

    Sample: brain
  10. JOURNAL OF MEDICINAL FOOD (2019) IF: 2.02
    Effects of Euterpe oleracea to Enhance Learning and Memory in a Conditioned Nicotinic and Muscarinic Receptor Response Paradigm by Modulation of Cholinergic Mechanisms in Rats

    DOI: 10.1089/jmf.2018.0197

    PMID: 31580752

    Sample: Tissue homogenate
  11. NEUROSCIENCE LETTERS (2019) IF: 2.173
    Differentiation of human glioblastoma U87 cells into cholinergic neuron

    DOI: 10.1016/j.neulet.2019.03.049

    PMID: 30928478

    Sample: Cell culture supernatant
  12. NEUROLOGICAL RESEARCH (2023) IF: 2.529
    The effect of the calcium channel blocker nimodipine on hippocampal BDNF/Ach levels in rats with experimental cognitive impairment

    DOI: 10.1080/01616412.2022.2164452

    Sample: Hippocampal,prefrontal cortex
  13. Evidence-based Complementary and Alternative Medicine (2021) IF: 2.63
    Standardized Extract (HemoHIM) Protects against Scopolamine-Induced Amnesia in a Murine Model

    DOI: 10.1155/2021/8884243

    PMID: 33815562

    Sample: serum
  14. Journal of General and Family Medicine (2023)
    The role of biofield energy treatment on psychological symptoms, mental health disorders, and stress-related quality of life in adult subjects: A randomized controlled clinical trial

    DOI: 10.1002/jgf2.606

    Sample: serum
  15. Veterinary World (2018)
    The analysis of hippocampus neuronal density (CA1 and CA3) after Ocimum sanctum ethanolic extract treatment on the young adulthood and middle age rat model

    DOI: 10.14202/vetworld.2018.135-140

    Sample: Brain tissues
  16. Advances in Traditional Medicine (2022)
    Nootropic effect of Syzygium polyanthum (Wight) Walp leaf extract in chronic cerebral hypoperfusion rat model via cholinergic restoration: a potential therapeutic agent for dementia

    DOI: 10.1007/s13596-022-00653-3

    Sample: brain
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