ELISA Kit for Vasoactive Intestinal Peptide (VIP)

PHM27; Intestinal peptide PHV-42; Peptide histidine valine 42; Peptide histidine methioninamide 27

Specificity

This assay has high sensitivity and excellent specificity for detection of Vasoactive Intestinal Peptide (VIP).
No significant cross-reactivity or interference between Vasoactive Intestinal Peptide (VIP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Vasoactive Intestinal Peptide (VIP) and the recovery rates were calculated by comparing the measured value to the expected amount of Vasoactive Intestinal Peptide (VIP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-101 89
EDTA plasma(n=5) 80-101 91
heparin plasma(n=5) 86-94 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vasoactive Intestinal Peptide (VIP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vasoactive Intestinal Peptide (VIP) 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 Vasoactive Intestinal Peptide (VIP) 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) 78-102% 91-98% 81-88% 90-104%
EDTA plasma(n=5) 93-101% 78-103% 90-97% 80-93%
heparin plasma(n=5) 87-94% 99-105% 83-103% 80-104%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Clinical Genetics VIP gene variants related to idiopathic pulmonary arterial hypertension in Chinese population Wiley: source
Human immunology Low levels of vasoactive intestinal peptide are associated with Chagas disease cardiomyopathy Pubmed: 23800435
Headache: The Journal of Head and Face Pain CGRP and VIP Levels as Predictors of Efficacy of Onabotulinumtoxin Type A in Chronic Migraine Onlinelibrary: head.12372
World J Gastroenterol Roles of sphincter of Oddi motility and serum vasoactive intestinal peptide, gastrin and cholecystokinin octapeptide NCBI: PMC4000510
Cephalalgia? Increased VIP levels in peripheral blood outside migraine attacks as a potential biomarker of cranial parasympathetic activation in chronic migraine Pubmed:24847167
J Huazhong Univ Sci Technolog Med Sci. Clinical study of electro-acupuncture treatment with different intensities for functional constipation patients Pubmed:25318892
PLOS ONE Aqueous Fraction of Beta vulgaris Ameliorates Hyperglycemia in Diabetic Mice due to Enhanced Glucose Stimulated Insulin Secretion, Mediated by Acetylcholine and GLP-1, and Elevated Glucose Uptake via Increased Membrane Bound GLUT4 Transporters Pubmed:25647228
PLOS ONE Aqueous Fraction of PubMed: 25647228
Pharmacol Res Oral curcumin has anti-arthritic efficacy through somatostatin generation PubMed: 25836921
World J Gastroenterol Effects of sphincter of Oddi motility on the formation of cholesterol gallstones Pubmed:27350732
Annals of Allergy, Asthma and Immunology Effect of acupuncture on house dust mite specific IgE, substance P, and symptoms in persistent allergic rhinitis Pubmed:27156748
Cephalalgia. Relationship between serum levels of VIP, but not of CGRP, and cranial autonomic parasympathetic symptoms: A study in chronic migraine patients Pubmed:27250233
Headache No Change in Interictal PACAP Levels in Peripheral Blood in Women With Chronic Migraine pubmed:27634731
Sci Rep. Using the "target constituent removal combined with bioactivity assay" strategy to investigate the optimum arecoline content in charred areca nut. pubmed:28054652
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