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SP(Substance P) ELISA Kit

Cat.No.:E-EL-0067
Reactivity: Universal

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96T
48T
24T
96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	78.13-5000 pg/mL
Sensitivity	46.88 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes SP in samples. No significant cross-reactivity or interference between SP 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 SP 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 SP. During the reaction, SP in the sample or standard competes with a fixed amount of SP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to SP. 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 SP 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 SP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level SP 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)	233.17	510.94	2017.45	206.77	515.17	2371.94
Standard deviation	12.87	23.61	104.91	12.18	23.85	125.24
CV (%)	5.52	4.62	5.20	5.89	4.63	5.28

Recovery

The recovery of SP 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)	86-96	93
EDTA plasma(n=8)	99-109	104
Cell culture media(n=8)	90-100	96

Target Information

Research Area

Neuroscience, Cardiovascular

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

BASIC RESEARCH IN CARDIOLOGY (2022) IF: 12.416
Spinal cord astrocytes regulate myocardial ischemia–reperfusion injury

DOI: 10.1007/s00395-022-00968-x

Sample: serum
Nutrients (2022) IF: 6.706
Prevention of Loperamide-Induced Constipation in Mice and Alteration of 5-Hydroxytryotamine Signaling by Ligilactobacillus salivarius Li01

DOI: 10.3390/nu14194083

PMID: 36235735
Sample: serum
PHYTOMEDICINE (2021) IF: 5.34
Extracts of Poria cocos improve functional dyspepsia via regulating brain-gut peptides, immunity and repairing of gastrointestinal mucosa

DOI: 10.1016/j.phymed.2021.153875

PMID: 34911003
Sample: serum
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
JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
Protective effect of the combination of essential oil from patchouli and tangerine peel against gastric ulcer in rats

DOI: 10.1016/j.jep.2021.114645

PMID: 34530094
Sample: serum
JOURNAL OF ETHNOPHARMACOLOGY (2020) IF: 4.36
Laxative effect and mechanism of Tiantian Capsule on loperamide-induced constipation in rats

DOI: 10.1016/j.jep.2020.113411

PMID: 32980482
Sample: Serum
JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
Atractylodes oil alleviates diarrhea-predominant irritable bowel syndrome by regulating intestinal inflammation and intestinal barrier via SCF/c-kit and MLCK/MLC2 pathways

DOI: 10.1016/j.jep.2021.113925

PMID: 33592255
Sample: Serum,Tissue homogenate(colon)
Aging-US (2020) IF: 4.831
NO up-regulates migraine-related CGRP via activation of an Akt/GSK-3β/NF-κB signaling cascade in trigeminal ganglion neurons

DOI: 10.18632/aging.103031

Sample: Primary TGNs
Frontiers in Medicine (2020) IF: 3.9
Acute Effect of Transcutaneous Electroacupuncture on Globus Pharyngeus: A Randomized, Single-Blind, Crossover Trial

DOI: 10.3389/fmed.2020.00179

PMID: 32528966
Sample: Plasma
NEUROSCIENCE LETTERS (2020) IF: 2.274
FSC231 can alleviate paclitaxel-induced neuralgia by inhibiting PICK1 and affecting related factors

DOI: 10.1016/j.neulet.2020.135471

PMID: 33207243
Sample: Tissue homogenate
BRAIN RESEARCH (2019) IF: 2.929
Cervical sympathectomy modulates the neurogenic inflammatory neuropeptides following experimental subarachnoid hemorrhage in rats

DOI: 10.1016/j.brainres.2019.146366

PMID: 31401069
Sample: Plasma,Tissue homogenate
ORL-Journal for Oto-Rhino-Laryngology Head and Neck Surgery (2021) IF: 1.538
Clinical Efficacy and Possible Mechanism of Endoscopic Vidian Neurectomy for House Dust Mite-Sensitive Allergic Rhinitis

DOI: 10.1159/000511711

Sample: serum
Evidence-based Complementary and Alternative Medicine (2020) IF: 1.813
Laxative Effects of Yangyin Tongmi Capsule on a Model of Diphenoxylate-Induced Constipation in Mice

DOI: 10.1155/2020/1471824

PMID: 32148532
Sample: Serum
Revista da Associacao Medica Brasileira (2019) IF: 0.801
Modulatory effects of neuropeptides on pentylenetetrazol-induced epileptic seizures and neuroinflammation in rats

DOI: 10.1590/1806-9282.65.9.1188

PMID: 31618336
Sample: Plasma

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