CLIA Kit for Substance P (SP) Mus musculus (Mouse) Competition CLIA

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  • CLIA Kit for Substance P (SP) Packages (Simulation)
  • CLIA Kit for Substance P (SP) Packages (Simulation)
  • CLIA Kit for Substance P (SP) Results demonstration
  • CCA393Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 97-104 101
EDTA plasma(n=5) 80-101 98
heparin plasma(n=5) 93-101 98

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) 78-101% 94-101% 94-105% 95-102%
EDTA plasma(n=5) 88-103% 80-104% 82-101% 83-96%
heparin plasma(n=5) 90-101% 82-98% 99-105% 95-105%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
7. Read RLU value immediately.

CLIA Kit for Substance P (SP)

Test principle

The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Substance P (SP). A competitive inhibition reaction is launched between biotin labeled Substance P (SP) and unlabeled Substance P (SP) (Standards or samples) with the pre-coated antibody specific to Substance P (SP). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Substance P (SP) in the sample. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is reverse proportional to the Substance P (SP) level in the sample or standard.

Citations

  • A Potential Role for Substance P and Interleukin-6 in the Cerebrospinal Fluid of Cavalier King Charles Spaniels with Neuropathic PainPubmed: 23659719
  • Oral curcumin has anti-arthritic efficacy through somatostatin generationPubMed: 25836921
  • Positive enhancement of Lactobacillus fermentum HY01 on intestinal movements of mice having constipation10.1007/s13765-017-0327-3
  • Direct activation of tachykinin receptors within baroreflex afferent pathway and neurocontrol of blood pressure regulationPubmed:29900692
  • Serum Levels of Neuropeptides in Cows with Left Abomasal DisplacementPubmed: 30562932
  • Effects of nanoparticles on Neuroinflammation in a Mouse Model of AsthmaPubmed: 31542455
  • Hejie Zhitong prescription promotes sleep and inhibits nociceptive transmission-associated neurotransmitter activity in a rodent migraine modelPubmed: 33014123
  • Changes of substance P, NGF and CGRP salivary levels among patients undergoing physical therapy
  • Evaluation of the causes affecting the development of pruritus in patients with peritoneal dialysis34213714
  • Surfactant protein C is associated with perineuronal nets and shows age-dependent changes of brain content and hippocampal deposits in wildtype and 3xTg mice34626771

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