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Multiplex Assay Kit for Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay)

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 96
EDTA plasma(n=5) 80-97 86
heparin plasma(n=5) 80-94 87
sodium citrate plasma(n=5) 92-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Substance P (SP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 93-101% 90-98% 78-96% 81-90%
EDTA plasma(n=5) 92-101% 97-104% 91-103% 78-98%
heparin plasma(n=5) 96-104% 98-105% 78-104% 90-103%
sodium citrate plasma(n=5) 84-92% 78-104% 83-104% 86-94%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:SP) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Journal of Veterinary Internal Medicine A Potential Role for Substance P and Interleukin-6 in the Cerebrospinal Fluid of Cavalier King Charles Spaniels with Neuropathic Pain Pubmed: 23659719
Pharmacol Res Oral curcumin has anti-arthritic efficacy through somatostatin generation PubMed: 25836921
Applied Biological Chemistry Positive enhancement of Lactobacillus fermentum HY01 on intestinal movements of mice having constipation 10.1007/s13765-017-0327-3
CNS Neuroscience & Therapeutics  Direct activation of tachykinin receptors within baroreflex afferent pathway and neurocontrol of blood pressure regulation Pubmed:29900692
Journal of Veterinary Science Serum Levels of Neuropeptides in Cows with Left Abomasal Displacement Pubmed: 30562932
Respiratory Physiology & Neurobiology Effects of nanoparticles on Neuroinflammation in a Mouse Model of Asthma Pubmed: 31542455
Chinese Medicine Hejie Zhitong prescription promotes sleep and inhibits nociceptive transmission-associated neurotransmitter activity in a rodent migraine model Pubmed: 33014123
Medical Research Journal Changes of substance P, NGF and CGRP salivary levels among patients undergoing physical therapy
Int Urol Nephrol Evaluation of the causes affecting the development of pruritus in patients with peritoneal dialysis 34213714
J Chem Neuroanat Surfactant protein C is associated with perineuronal nets and shows age-dependent changes of brain content and hippocampal deposits in wildtype and 3xTg mice 34626771
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