Rat PK2 (Prokineticin 2) CLIA Kit (RTES00470)

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ELISA Kit Technical ManualMSDS

Rat PK2 (Prokineticin 2) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat PK2 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat PK2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat PK2, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat PK2. The concentration of Rat PK2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Assay time4.5h
Detection methodChemiluminescence
Detection range62.50-4000 pg/mL
Sensitivity37.50 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%


This kit recognizes Rat PK2 in samples. No significant cross-reactivity or interference between Rat PK2 and analogues was observed.

Typical Data

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

RLU Average Corrected


Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat PK2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat PK2 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Mean (pg/mL)192.15624.951944.66205.01663.062018.56
Standard deviation16.0651.68197.7725.9555.70123.13
C V (%)8.368.2710.1712.668.406.10


The recovery of Rat PK2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)94-108100
EDTA plasma (n=5)84-9790
Cell culture media (n=5)85-9992


Samples were spiked with high concentrations of Rat PK2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)98-11597-11286-99
Average (%)10610592
1:4Range (%)95-10891-10685-98
Average (%)1019792
1:8Range (%)93-10597-11392-105
Average (%)98105100
1:16Range (%)96-11191-104104-117
Average (%)10499110

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro CLIA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Rat PK2 (Prokineticin 2) CLIA Kit (RTES00470) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Rat PK2 (Prokineticin 2) CLIA Kit (RTES00470) Protein Information

UniProt Protein Function:Function: May function as an output molecule from the suprachiasmatic nucleus (SCN) that transmits behavioral circadian rhythm. May also function locally within the SCN to synchronize output. Potently contracts gastrointestinal (GI) smooth muscle
UniProt Protein Details:

By similarity.

Subcellular location: Secreted

By similarity.

Tissue specificity: Expressed at high levels in testis and at lower levels in brain, lung, ovary, spleen, thymus and uterus.

Induction: Activated by CLOCK and BMAL1 heterodimers and light; inhibited by period genes (PER1, PER2 and PER3) and cryptochrome genes (CRY1 and CRY2)


Sequence similarities: Belongs to the AVIT (prokineticin) family.

NCBI Summary:ligand of of two orphan G-protein-coupled receptors designated ZAQ and I5E [RGD, Feb 2006]
UniProt Code:Q8R413
NCBI GenInfo Identifier:20301998
NCBI Gene ID:192206
NCBI Accession:NP_620207.1
UniProt Related Accession:Q8R413
Molecular Weight:10.2kDa
NCBI Full Name:prokineticin-2 isoform 2
NCBI Synonym Full Names:prokineticin 2
NCBI Official Symbol:Prok2  
NCBI Official Synonym Symbols:Bv8  
NCBI Protein Information:prokineticin-2; PK2; prokineticin 2 beta; homolog of mouse Bv8 (Bombina variegata 8 kDa); homolog of mouse Bv8 (Bombina variegata 8 kDa); prokineticin 2
UniProt Protein Name:Prokineticin-2
Protein Family:Probable serine/threonine-protein kinase
UniProt Gene Name:Prok2  
UniProt Entry Name:PROK2_RAT
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Additional Information

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