CLIA Kit for Calcitonin Gene Related Peptide (CGRP)

CALCB; CGRP2; CALC2; CGRP-II; Calcitonin Gene Related Peptide 2; Calcitonin 2; Calcitonin Related Polypeptide Beta

Specificity

This assay has high sensitivity and excellent specificity for detection of Calcitonin Gene Related Peptide (CGRP).
No significant cross-reactivity or interference between Calcitonin Gene Related Peptide (CGRP) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-98 85
EDTA plasma(n=5) 83-92 89
heparin plasma(n=5) 80-94 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Calcitonin Gene Related Peptide (CGRP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Calcitonin Gene Related Peptide (CGRP) 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 Calcitonin Gene Related Peptide (CGRP) 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) 80-102% 84-99% 89-97% 90-104%
EDTA plasma(n=5) 80-98% 91-99% 78-96% 78-101%
heparin plasma(n=5) 89-96% 98-105% 89-103% 88-96%

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 Assay Diluent A 1×12mL
Detection Reagent B 1×260µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Substrate B 1×2mL
Substrate A 1×10mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
7. Read RLU value immediately.

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Magazine Citations
Evidence-Based Complementary and Alternative Medicine Alstonia scholaris R. Br. Significantly Inhibits Retinoid-Induced Skin Irritation In Vitro and In Vivo. Hindawi: 190370
The Journal of Headache and Pain Increased levels of CGRP in peripheral blood in women with chronic migraine: A reliable biological marker PubMed: PMC3620076
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
PLoS One. Ultraviolet B Irradiation Reduces the Expression of Adiponectin in Ovarial Adipose Tissues through Endocrine Actions of Calcitonin Gene-Related Peptide-Induced Serum Amyloid A Pubmed:Pmc4028234
Int J Vasc Med. Prognostic significance of circulating and endothelial progenitor cell markers in type 2 diabetic foot. Pubmed:24624298
Arch Gynecol Obstet. Calcitonin gene-and parathyroid hormone-related peptides in normotensive and preeclamptic pregnancies: a nested case–control study Pubmed:24943060
Pain OnabotulinumtoxinA decreases interictal CGRP plasma levels in patients with chronic migraine PubMed: 25735000
Molecular Medicine Reports Exogenous α-calcitonin gene-related peptide attenuates lipopolysaccharide-induced acute lung injury in rats PubMed: 25892548
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
Cardiology.  Transient Receptor Potential Vanilloid Subtype 1 Inhibits Inflammation and Apoptosis via the Release of Calcitonin Gene-Related Peptide in the Heart after Myocardial Infarction. Pubmed:27144592
J Headache Pain Valproate ameliorates nitroglycerin-induced migraine in trigeminal nucleus caudalis in rats through inhibition of NF-кB Pubmed:27150105
Frontiers in Physiology Prevention of Bone Loss in a Model of Postmenopausal Osteoporosis throughAdrenomedullin Inhibition. pubmed:27445864
Naunyn Schmiedebergs Arch Pharmacol. Opioid receptor agonists may favorably affect bone mechanical properties in rats with estrogen deficiency-induced osteoporosis. pubmed:27896372
American Journal of Translational Research A new comorbidity model and the common pathological mechanisms of migraine and epilepsy pubmed:28559979
9 Probiotic Bifidobacterium longum NCC3001 Reduces Depression Scores and Alters Brain Activity: A Pilot Study in Patients With Irritable Bowel Syndrome pubmed:28483500
Journal of Clinical Investigation CNS myeloid cells critically regulate heat hyperalgesia Pubmed:29634489
Journal of Headache and Pain Feasibility of serum CGRP measurement as a biomarker of chronic migraine: a critical reappraisal Pubmed:30006780
BioRxiv - the preprint server for biology RAMP1 in Kupffer cells is a critical regulator in immune-mediated hepatitis 10.1101:357582
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