Human SOST/Sclerostin Quantikine ELISA Kit

R&D Systems | Catalog # DSST00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

3.8 pg/mL

Assay Range

31.3-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
View all SOST/Sclerostin ELISA Kits »

Product Summary for Human SOST/Sclerostin Quantikine ELISA Kit

The Quantikine Human SOST immunoassay is a 4.5 hour solid-phase ELISA designed to measure SOST in human cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human SOST and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant human SOST. Results obtained using natural human 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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human 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.

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. Assays were performed by at least three technicians using two lots of components.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 308 577 1208 315 591 1234
Standard Deviation 6.03 10.5 25.1 25.9 63.8 118
CV% 2.0 1.8 2.1 8.2 10.8 9.6

Recovery for Human SOST/Sclerostin Quantikine ELISA Kit

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 Media (n=4) 100 91-111
EDTA Plasma (n=4) 95 89-101
Heparin Plasma (n=4) 100 91-108
Serum (n=4) 96 86-112

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human SOST were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human SOST/Sclerostin ELISA Linearity

Scientific Data Images for Human SOST/Sclerostin Quantikine ELISA Kit

Human SOST/Sclerostin ELISA Standard Curve

Human SOST/Sclerostin ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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.

Alternate Names

sclerostin, VBCHsclerosteosis

Entrez Gene IDs

50964 (Human); 74499 (Mouse)

Gene Symbol

SOST

Additional SOST/Sclerostin Products

Product Documents for Human SOST/Sclerostin Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human SOST/Sclerostin Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human SOST/Sclerostin Quantikine ELISA Kit

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Protocols

View specific protocols for Human SOST/Sclerostin Quantikine ELISA Kit (DSST00):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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