< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Human SPARC Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human SPARC in cell culture supernates, serum, plasma, and human milk. It contains NS0-expressed recombinant human SPARC and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human SPARC showed linear 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 naturally occurring human SPARC.
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.
The recovery of SPARC spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
Platelet-poor EDTA Plasma (n=4)
Platelet-poor Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of SPARC were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
SPARC, also known as Osteonectin and Basement Membrane protein BM-40, is a phosphorylated glycoprotein with calcium-binding domains. It is secreted by endothelial cells and is present in large amounts in the parietal endoderm of mouse embryos and in human placenta. SPARC may be involved in cell proliferation, repair of tissue damage, and morphogenic processes such as re-modeling of the extracellular matrix.
Secreted Protein Acidic and Rich in Cysteine
Entrez Gene IDs
6678 (Human); 20692 (Mouse);
Basement-membrane protein 40; BM-40; ONcysteine-rich protein; osteonectin; Secreted protein acidic and rich in cysteine; secreted protein, acidic, cysteine-rich (osteonectin);
Refer to the product for complete assay procedure.
The conjugate must remain at 2-8 °C during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 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.
Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.
200 µL cold Conjugate
Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour on the benchtop.
Aspirate and wash 6 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
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.
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.