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Mouse β-EP(Beta-Endorphin) ELISA Kit

Citations(6)Uniprot : P01193

Cat.No.:E-EL-M0184
Reactivity: Mouse

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96T
48T
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96T*5
96T*10

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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 β-EP in samples. No significant cross-reactivity or interference between β-EP 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 β-EP 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 β-EP. During the reaction, β-EP in the sample or standard competes with a fixed amount of β-EP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to β-EP. 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 β-EP 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.

Item	Specifications	Storage
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 β-EP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level β-EP 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)	49.40	99.80	404.50	51.10	94.90	411.30
Standard deviation	2.90	5.50	14.20	3.20	5.40	14.80
CV (%)	5.87	5.51	3.51	6.26	5.69	3.60

Recovery

The recovery of β-EP 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)	89-105	96
EDTA plasma(n=8)	85-101	92
Cell culture media(n=8)	95-107	101

Linearity

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

Target Information

Database Links	SwissProt: P01193
Synonyms	b-EP
Research Area	Cancer, Metabolism, Neuroscience, Signal transduction

Assay Procedures

	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
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

Biomedicines (2021) IF: 6.081
Analgesic and Anti-Inflammatory Effects of the Synthetic Neurosteroid Analogue BNN27 during CFA-Induced Hyperalgesia

DOI: 10.3390/biomedicines9091185

PMID: 34572370
Sample: serum
NEUROPHARMACOLOGY (2017) IF: 5.012
Delta-opioid receptor antagonism leads to excessive ethanol consumption in mice with enhanced activity of the endogenous opioid system

DOI: 10.1016/j.neuropharm.2017.03.016

Sample: Plasma
MOLECULAR NEUROBIOLOGY (2018) IF: 5.076
Lymphocytes and Met-Enkephalin

DOI: 10.1007/s12035-018-1176-8

Sample: Plasma
NEUROPHARMACOLOGY (2018) IF: 4.249
Bidirectional selection for high and low stress-induced analgesia affects G-protein activity

DOI: 10.1016/j.neuropharm.2018.10.014

PMID: 30326238
Sample: Tissue homogenate
Molecular Pain (2018) IF: 3.205
Lithium reverses mechanical allodynia through a mu opioid-dependent mechanism:

DOI: 10.1177/1744806917754142

Sample: brain,plasma
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2016) IF: 2.371
G protein coupled receptor kinase-2 upregulation causes κ-opioid receptor desensitization in diabetic heart

DOI: 10.1016/j.bbrc.2016.11.090

Sample: Plasma

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