Mouse Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit

(1 citations)   
  • 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), Serum (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    2.37 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse Shh-N. This kit also recognizes low levels of Shh-N in rat serum.
  • 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 Sonic Hedgehog N-Terminus Immunoassay is a 4.5 hour solid phase ELISA designed to measure Shh-N in cell culture supernates, serum, and plasma. It contains recombinant mouse Shh-N and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse Shh-N showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse Shh-N.

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, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404444
Mean22.754.621221.950.4188.7
Standard Deviation1.081.3110.62.282.969.21
CV%4.82.4510.45.94.9

Recovery

The recovery of mouse Shh-N spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=6) 101 96-109
Heparin Plasma (n=4) 97 86-108
Serum (n=4) 98 86-109
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse Shh-N were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Sonic Hedgehog/Shh
Shh (Sonic Hedgehog) is expressed in embryonic tissues that are critical for the patterning of the developing central nervous system, somite, and limb. It is also involved in whisker, hair, foregut, tooth, and bone development. Shh regulates neural and hematopoietic stem cell fate and is important for thymocyte differentiation and proliferation. In adult tissue Shh is associated with cancer development and tissue remodeling following injury. Palmitoylation and cholesterol addition of Shh contribute to its membrane tethering and assembly into multimers. Lipid modification and multimerization greatly increase Shh signaling potency. Shh can be released from the plasma membrane by the cooperative action of DISP1, SCUBE2, and TACE/ADAM17. Canonical signaling of Shh is mediated by a receptor complex that includes Patched (PTCH1, PTCH2) and Smoothened (SMO).
    • Entrez Gene IDs
      6469 (Human); 20423 (Mouse);
    • Alternate Names
      HHG1; HHG-1; HLP3; HPE3; MCOPCB5sonic hedgehog (Drosophila) homolog; Shh; SMMCIsonic hedgehog homolog (Drosophila); sonic hedgehog homolog; sonic hedgehog protein; sonic hedgehog; TPT; TPTPS;
    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 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 2 hours.
    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. 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.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    Showing Results 1 - 1 of 1

    1. Spatially heterogeneous choroid plexus transcriptomes encode positional identity and contribute to regional CSF production.
      Authors: Lun M, Johnson M, Broadbelt K, Watanabe M, Kang Y, Chau K, Springel M, Malesz A, Sousa A, Pletikos M, Adelita T, Calicchio M, Zhang Y, Holtzman M, Lidov H, Sestan N, Steen H, Monuki E, Lehtinen M
      J Neurosci, 2015;35(12):4903-16.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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