Mouse/Rat WISP-1/CCN4 Quantikine ELISA Kit

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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL), Urine (50 uL)
  • Sensitivity
    2.74 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant mouse and rat WISP-1/CCN4
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse/Rat WISP-1/CCN4 immunoassay is a 4.5 hour solid-phase ELISA designed to measure WISP-1 in mouse or rat cell culture supernates, tissue lysates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse WISP-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate
the recombinant mouse WISP-1. Results obtained using natural mouse or rat WISP-1 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural WISP-1.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components.
Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean33.810632337.393.7288
Standard Deviation1.584.0615.93.596.1611.1
CV%4.73.84.99.66.63.9

Recovery

The recovery of WISP-1 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 104 89-119
Cell Lysates (n=4) 89 80-111
EDTA Plasma (n=4) 110 96-119
Heparin Plasma (n=4) 104 81-120
Serum (n=4) 104 90-119
Urine (n=4) 107 90-120
Linearity
To assess the linearity of the assay, samples containing high concentrations of WISP-1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse/Rat WISP-1/CCN4 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: WISP-1/CCN4
WISP-1 belongs to the CCN (CTGF/CYR61/NOV) family of secreted proteins that share a common multimodular organization. Each family member has an IGFBP domain, a von Willebrand type C domain, a thrombospondin 1 domain and a C-terminal cysteine knot domain. Depending on cell context, WISP-1 can either promote or inhibit tumorigenesis.
    • Long Name
      Wnt-1 Induced Secreted Protein 1
    • Entrez Gene IDs
      8840 (Human); 22402 (Mouse);
    • Alternate Names
      CCN family member 4; CCN4; CCN4WISP1tc; WISP1; WISP-1; WISP1c; WISP1i; WNT1 induced secreted protein 1; WNT1 inducible signaling pathway protein 1; Wnt-1 inducible signaling pathway protein 1; wnt-1 signaling pathway protein 1; Wnt-1-induced secreted protein; WNT1-inducible-signaling pathway protein 1;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 4 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Cell Lysis Buffer 2

    ELISA 895347 2
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    Average Rating: 5 (Based on 1 review)

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     Mouse/Rat WISP-1/CCN4 Quantikine ELISA Kit MWSP10
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      Anonymous 07/12/2017
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     Mouse/Rat WISP-1/CCN4 Quantikine ELISA Kit MWSP10
    : Mouse/Rat WISP-1/CCN4 Quantikine ELISA Kit [MWSP10]

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