Human SPARC Quantikine ELISA Kit

R&D Systems | Catalog # DSP00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (10 µL), Serum (10 µL), Platelet-poor EDTA Plasma (20 µL), Platelet-poor Heparin Plasma (20 µL), Human Milk (50 µL)

Sensitivity

0.269 ng/mL

Assay Range

1.56-50 ng/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Human Milk)
View all SPARC ELISA Kits »

Product Summary for Human SPARC Quantikine ELISA Kit

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.

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 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.

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, Human Milk, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 7.34 14.3 29.9 6.65 13.5 28.3
Standard Deviation 0.129 0.282 0.733 0.541 0.989 2.40
CV% 1.8 2.0 2.5 8.1 7.3 8.5

Recovery for Human SPARC Quantikine ELISA Kit

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.

Human SPARC ELISA Linearity

Scientific Data Images for Human SPARC Quantikine ELISA Kit

Human SPARC ELISA Standard Curve

Human SPARC 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: 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

Alternate Names

BM-40, Osteonectin

Entrez Gene IDs

6678 (Human); 20692 (Mouse)

Gene Symbol

SPARC

Additional SPARC Products

Product Documents for Human SPARC 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 SPARC 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 SPARC Quantikine ELISA Kit

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  • Human SPARC Quantikine ELISA Kit
    Name: Denis Streltsov
    Sample Tested: Cell culture supernatant and Tissue Culture Media
    Verified Customer | Posted 11/21/2018
    Human SPARC Quantikine ELISA Kit DSP00

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Protocols

View specific protocols for Human SPARC Quantikine ELISA Kit (DSP00):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.
    6. 200 µL cold Conjugate
  6.   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.
  7.   Aspirate and wash 6 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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