Mouse Pentraxin 2/SAP Quantikine ELISA Kit

  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (10 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.368 ng/mL
  • Assay Range
    1.6 - 100 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse PTX2/SAP
  • 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 PTX2/SAP Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse PTX2/SAP in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse PTX2/SAP and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse PTX2/SAP showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse PTX2/SAP.

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 known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean4.111633.63.9515.734.5
Standard Deviation0.220.571.330.330.761.59
CV%5.43.648.44.84.6

Recovery

The recovery of PTX2/SAP spiked into cell culture samples was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=6) 97 81-112
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse PTX2/SAP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse Pentraxin 2/SAP Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Pentraxin 2/SAP
Pentraxin or Pentraxin 2 (PTX2), also known as Serum Amyloid P component (SAP), is a secreted serum glycoprotein belonging to the pentraxin superfamily. Pentraxins have a characteristic pentagonal discoid arrangement of five non-covalently bound subunits and bind a variety of molecules in a calcium- dependent lectin-like manner through a pattern-recognition-binding site. PTX2 is a major acute-phase protein and may function as an opsonin for a variety of ligands. PTX2 also associates ubiquitously with all amyloid deposits implicated in diseases including Alzheimer's and prion diseases, type 2 diabetes and various systemic amyloidoses.
    • Long Name
      Serum Amyloid P Component
    • Entrez Gene IDs
      325 (Human); 20219 (Mouse); 29339 (Rat);
    • Alternate Names
      9.5S alpha-1-glycoprotein; amyloid P component, serum; APCS; MGC88159; Pentraxin 2; PTX2; PTX2serum amyloid P-component; SAP; SAPpentaxin-related; Serum Amyloid P component;
    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 Calibrator Diluent
    4.   Add 50 µL of Calibrator 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 3 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 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 1 hour on the shaker.
    10.   Aspirate and wash 5 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. 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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