Human Glypican 3 Quantikine ELISA Kit

  
  • 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 (100 uL), Cell Lysates (20 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
  • Sensitivity
    20.6 pg/mL
  • Assay Range
    78.1 - 5,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human Glypican 3.
  • 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.
Control Available
QC203, Quantikine Immunoassay Control Set 848 for Human Glypican - Please Inquire
Product Summary
The Quantikine Human Glypican 3 Immunoassay is a 4.5 hour solid phase ELISA designed to measure Glypican 3 levels in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant human Glypican 3 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human Glypican 3 showed linear curves that were parallel to the standard curves obtained using the Quantikine Human Glypican 3 Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human Glypican 3.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean4121208238746612312359
Standard Deviation124879.638.475.3143
CV%2.943.38.26.16.1

Recovery

The recovery of human Glypican 3 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 88-112
EDTA Plasma (n=4) 88 81-95
Heparin Plasma (n=4) 89 81-96
Serum (n=4) 89 82-97
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Glypican 3 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Glypican 3 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Glypican 3

The glypicans (GPC) constitute a family of glycosylphosphatidylinositol (GPI)-anchored, heparan sulfate proteoglycans. Six members of this family have been identified in mammals (GPC1-GPC6). All glypican core proteins contain an N-terminal signal peptide, a large globular cysteine-rich domain (CRD) with 14 invariant cysteine residues, a stalk-like region containing the heparan sulfate attachment sites, and a C-terminal GPI attachment site.

Mutations in Glypican 3 cause a rare disorder in humans, Simpson-Golabi-Behmel Syndrome, which is characterized by pre- and postnatal overgrowth of multiple tissues and organs and an increased risk for developing embryonic tumors. These features are also present in the mouse knock-out of Glypican 3 indicating that Glypican 3 regulates cell survival and inhibits cell proliferation during development. Glypican 3 has been implicated in regulating many different signaling pathways including: IGF, FGF, BMP and Wnt. An endoproteolytic processing of Glypican 3 by proprotein convertases is required for the modulation of Wnt signaling. Direct interaction of Glypican 3 with FGF-basic has been observed and is mediated by the heparan sulfate chains.

    • Entrez Gene IDs
      2719 (Human); 14734 (Mouse); 25236 (Rat);
    • Alternate Names
      DGSX; Glypican 3; glypican proteoglycan 3; glypican-3; GPC3; GTR2-2; heparan sulphate proteoglycan; Intestinal protein OCI-5; MXR7; OCI5; OCI-5; secreted glypican-3; SGB; SGBS; SGBS1SDYS;
    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. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 848 for Human Glypican

    Ctrl QC203
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Cell Lysis Buffer 1 (1 x 21 mL)

    ELISA 890713 1
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