Human SPARC Quantikine ELISA Kit

  (3 citations)     
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Product Details
Assay Procedure
Citations (3)
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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 (10 uL), Serum (10 uL), Platelet-poor EDTA Plasma (20 uL), Platelet-poor Heparin Plasma (20 uL), Human Milk (50 uL)
  • Sensitivity
    0.269 ng/mL
  • Assay Range
    1.6 - 50 ng/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Human Milk)
  • Specificity
    Natural and recombinant human SPARC
  • 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
QC160 , Quantikine Immunoassay Control Set 659 for Human SPARC - Please Inquire
Product Summary
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.

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, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Human Milk
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 7.34 14.3 29.9 6.65 13.5 28.3
Standard Deviation 0.129 0.282 0.733 0.541 0.989 2.4
CV% 1.8 2 2.5 8.1 7.3 8.5

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 86-111
Platelet-poor EDTA Plasma (n=4) 104 93-111
Platelet-poor Heparin Plasma (n=4) 91 82-100
Linearity
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.
 SPARC [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: SPARC
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.
  • Long Name:
    Secreted Protein Acidic and Rich in Cysteine
  • Entrez Gene IDs:
    6678 (Human); 20692 (Mouse)
  • Alternate Names:
    Basement-membrane protein 40; BM-40; ONcysteine-rich protein; Osteonectin; Secreted protein acidic and rich in cysteine; secreted protein, acidic, cysteine-rich (osteonectin); SPARC
Related Research Areas
Assay Procedure
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.
  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 3 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.

  8. 200 µL cold Conjugate
  9.   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.
  10.   Aspirate and wash 6 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.
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.

3 Citations: Showing 1 - 3
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Species
Sample Type
  1. SPARC expression is associated with hepatic injury in rodents and humans with non-alcoholic fatty liver disease
    Authors: G Mazzolini, C Atorrasaga, A Onorato, E Peixoto, M Schlattjan, JP Sowa, S Sydor, G Gerken, A Canbay
    Sci Rep, 2018;8(1):725.
    Species: Human
    Sample Type: Serum
  2. SPARC Expression Did Not Predict Efficacy of nab-Paclitaxel plus Gemcitabine or Gemcitabine Alone for Metastatic Pancreatic Cancer in an Exploratory Analysis of the Phase III MPACT Trial.
    Authors: Hidalgo M, Plaza C, Musteanu M, Illei P, Brachmann C, Heise C, Pierce D, Lopez-Casas P, Menendez C, Tabernero J, Romano A, Wei X, Lopez-Rios F, Von Hoff D
    Clin Cancer Res, 2015;21(21):4811-8.
    Species: Human
    Sample Type: Plasma
  3. SPARC expression in CML is associated to imatinib treatment and to inhibition of leukemia cell proliferation.
    Authors: Giallongo C, La Cava P, Tibullo D, Barbagallo I, Parrinello N, Cupri A, Stagno F, Consoli C, Chiarenza A, Palumbo G, Di Raimondo F
    BMC Cancer, 2013;13(0):60.
    Species: Human
    Sample Type: Serum

FAQs

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ELISA Controls
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Quantikine Immunoassay Control Set 659 for Human SPARC

Ctrl QC160
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Supplemental ELISA Products
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Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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