ELISA Kit for Pituitary Adenylate Cyclase Activating Peptide (PACAP) Homo sapiens (Human) Competition ELISA

ADCYAP1; PRP-48; Pituitary Adenylate Cyclase Activating Peptide

Add to Cart Distributors
Overview
Properties
Share your citation Upload your experimental result Review Leave a message
Loading...

Share a new citation as an author

Upload your experimental result

Review

Please attach serial No. on instruction manual

Contact us

Please fill in the blank.

Name*
Organization
Address
E-mail address*
Telephone
Inquiry*
Verification code* CheckCode
  • ELISA Kit for Pituitary Adenylate Cyclase Activating Peptide (PACAP) Packages (Simulation)
  • ELISA Kit for Pituitary Adenylate Cyclase Activating Peptide (PACAP) Packages (Simulation)
  • ELISA Kit for Pituitary Adenylate Cyclase Activating Peptide (PACAP) Results demonstration
  • CEB347Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Pituitary Adenylate Cyclase Activating Peptide (PACAP) and the recovery rates were calculated by comparing the measured value to the expected amount of Pituitary Adenylate Cyclase Activating Peptide (PACAP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-104 101
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pituitary Adenylate Cyclase Activating Peptide (PACAP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pituitary Adenylate Cyclase Activating Peptide (PACAP) 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 Pituitary Adenylate Cyclase Activating Peptide (PACAP) 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) 85-99% 96-104% 90-103% 93-102%
EDTA plasma(n=5) 91-99% 80-91% 83-90% 80-103%
heparin plasma(n=5) 84-93% 78-96% 93-101% 82-94%

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
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.

ELISA Kit for Pituitary Adenylate Cyclase Activating Peptide (PACAP)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Pituitary Adenylate Cyclase Activating Peptide (PACAP) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Pituitary Adenylate Cyclase Activating Peptide (PACAP) and unlabeled Pituitary Adenylate Cyclase Activating Peptide (PACAP) (Standards or samples) with the pre-coated antibody specific to Pituitary Adenylate Cyclase Activating Peptide (PACAP). 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 Pituitary Adenylate Cyclase Activating Peptide (PACAP) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Pituitary Adenylate Cyclase Activating Peptide (PACAP) in the sample.

Citations

  • Interictal plasma pituitary adenylate cyclase-activating polypeptide levels are decreased in migraineurs but remain unchanged in patients with tension-type headachePubMed: 26296898
  • Significance of serum VIP and PACAP in multiple sclerosis: an exploratory case–control study34698942

Recommend products