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ELISA Kit for Substance P (SP)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 82-105 86
EDTA plasma(n=5) 80-91 86
heparin plasma(n=5) 80-95 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Substance P (SP) 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) 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) 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) 83-102% 78-89% 86-95% 79-89%
EDTA plasma(n=5) 91-105% 95-104% 97-104% 96-104%
heparin plasma(n=5) 81-96% 87-94% 82-103% 80-102%

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
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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
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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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