Mouse/Rat SOST/Sclerostin Quantikine ELISA Kit

  (5 citations)   
Product Details
Assay Procedure
Citations (5)
FAQs
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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), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
  • Sensitivity
    4.17 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL
  • Specificity
    Natural and recombinant mouse and rat SOST
  • 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 SOST immunoassay is a 4.5 hour solid-phase ELISA designed to measure SOST in mouse or rat cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse SOST and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse SOST. Results obtained using natural mouse or rat SOST showed dose response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural SOST.

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 57.6 108 345 54.4 101 347
Standard Deviation 4.15 6 18.7 4.35 6.23 19.4
CV% 7.2 5.6 5.4 8 6.2 5.6

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 101 92-112
Mouse EDTA Plasma (n=4) 98 82-113
Mouse Heparin Plasma (n=4) 102 88-115
Mouse Serum (n=4) 95 86-113
Rat EDTA Plasma (n=2) 107 101-118
Rat Heparin Plasma (n=2) 96 82-111
Rat Serum (n=4) 99 85-116
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of SOST in each matrix were diluted with Calibrator Diluent and assayed.
Mouse/Rat SOST/Sclerostin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: SOST/Sclerostin
SOST, also known as Sclerostin, is a Cerberus/DAN family BMP antagonist and is an important regulator of bone homeostasis. Inactivating mutations in the SOST gene can cause sclerosteosis and van Buchem disease. SOST is expressed by terminally differentiated cells embedded in mineralized matrix including osteocytes, hypertrophic and prehypertrophic chondrocytes, and tooth cementocytes. Its expression is induced by BMP-2, -4, and -6 and is inhibited by parathyroid hormone (PTH). Its downregulation in osteocytes by physical loading of bone contributes to the mechanical sensor function of osteocytes and the subsequent increase in bone growth. SOST binds to BMP-2, -4, -5, -6, and -7 and inhibits the osteogenic differentiation induced by these BMPs. It inhibits canonical Wnt signaling by binding to LRP-5 and LRP-6 and inhibiting their association with Frizzled receptors. SOST reduces the proliferation of mesenchymal stem cells (MSC) and induces MSC apoptosis. Circulating levels of SOST are elevated in pathologies with bone involvement including low bone mineral density, Paget’s disease, multiple myeloma, and prostate cancer with bone metastases. It is also elevated in advanced chronic kidney disease, alcoholic liver disease, type 2 diabetes, and patients with increased abdominal and gynoid fat.
  • Entrez Gene IDs:
    50964 (Human); 74499 (Mouse)
  • Alternate Names:
    sclerostin; SOST; VBCHsclerosteosis
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, control, 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 3 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 1 hour.
  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.

5 Citations: Showing 1 - 5
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Species
Sample Type
  1. Acute Phosphate Restriction Impairs Bone Formation and Increases Marrow Adipose Tissue in Growing Mice
    Authors: Frank C Ko
    J Bone Miner Res, 2016;0(0):.
    Species: Mouse
    Sample Type: Serum
  2. Short-term intermittent administration of parathyroid hormone facilitates osteogenesis by different mechanisms in cancellous and cortical bone
    Authors: K Ogura, T Iimura, Y Makino, A Sugie-Oya, A Takakura, R Takao-Kawa, T Ishizuya, K Moriyama, A Yamaguchi
    Bone Rep, 2016;5(0):7-14.
    Species: Rat
    Sample Type: Serum
  3. Sclerostin inhibition prevents spinal cord injury-induced cancellous bone loss.
    Authors: Beggs L, Ye F, Ghosh P, Beck D, Conover C, Balaez A, Miller J, Phillips E, Zheng N, Williams A, Aguirre J, Wronski T, Bose P, Borst S, Yarrow J
    J Bone Miner Res, 2015;30(4):681-9.
    Species: Rat
    Sample Type: Serum
  4. Androgens inhibit the osteogenic response to mechanical loading in adult male mice.
    Authors: Sinnesael M, Laurent M, Jardi F, Dubois V, Deboel L, Delisser P, Behets G, D'Haese P, Carmeliet G, Claessens F, Vanderschueren D
    Endocrinology, 2015;156(4):1343-53.
    Species: Mouse
    Sample Type: Serum
  5. Role of TGF-beta in a mouse model of high turnover renal osteodystrophy.
    Authors: Liu S, Song W, Boulanger J, Tang W, Sabbagh Y, Kelley B, Gotschall R, Ryan S, Phillips L, Malley K, Cao X, Xia T, Zhen G, Cao X, Ling H, Dechow P, Bellido T, Ledbetter S, Schiavi S
    J Bone Miner Res, 2014;29(5):1141-57.
    Species: Mouse
    Sample Type: Serum

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