Human SPARC ELISA Kit (ab220654)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 115 pg/ml
- Range: 125 pg/ml - 8000 pg/ml
- Sample type: Cell culture media, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human SPARC ELISA Kit
See all SPARC kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 8 2.3% Inter-assay Sample n Mean SD CV% Serum 3 4.5% -
Sample type
Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
115 pg/ml -
Range
125 pg/ml - 8000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 125 115% - 130% Cell culture media 86 83% - 91% Hep Plasma 126 119% - 131% EDTA Plasma 124 115% - 132% Cit plasma 113 110% - 114% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Predicted to work with: Cow
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Product overview
Human SPARC ELISA Kit (ab220654) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of SPARC protein in cell culture media, cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human SPARC with 115 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
SPARC is a 286-amino acid, 43 kDa protein with two calcium binding sites as well as the ability to bind copper, collagen, albumin, and cell membranes. Fibroblasts, capillary endothelial cells, platelets, macrophages, and adipocytes all produce SPARC protein. Additionally, SPARC is involved in cell proliferation, repair of tissue damage, collagen matrix formation, and osteoblast differentiation. Importantly, because SPARC is present in platelet granules and is released upon platelet activation, to properly assess circulating levels of SPARC in plasma it is recommended to prepare and analyze platelet-poor plasma samples. Mouse, rat, and bovine SPARC are 92.4%, 92.4%, and 99.0% identical to human SPARC, respectively.
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Platform
Microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human SPARC Detector Antibody 1 x 600µl 10X Human SPARC Capture Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Human SPARC Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Appears to regulate cell growth through interactions with the extracellular matrix and cytokines. Binds calcium and copper, several types of collagen, albumin, thrombospondin, PDGF and cell membranes. There are two calcium binding sites; an acidic domain that binds 5 to 8 Ca(2+) with a low affinity and an EF-hand loop that binds a Ca(2+) ion with a high affinity. -
Sequence similarities
Belongs to the SPARC family.
Contains 1 EF-hand domain.
Contains 1 follistatin-like domain.
Contains 1 Kazal-like domain. -
Developmental stage
Expressed at high levels in tissues undergoing morphogenesis, remodeling and wound repair. -
Cellular localization
Secreted > extracellular space > extracellular matrix > basement membrane. In or around the basement membrane. - Information by UniProt
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Alternative names
- AA517111
- Basement membrane protein 40
- Basement-membrane protein 40
see all -
Database links
- Entrez Gene: 282077 Cow
- Entrez Gene: 6678 Human
- Omim: 182120 Human
- SwissProt: P13213 Cow
- SwissProt: P09486 Human
- Unigene: 111779 Human
- Unigene: 708558 Human
Images
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Other - Human SPARC ELISA Kit (ab220654)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human SPARC standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native SPARC in human serum and plasma samples.The concentration of SPARC was measured in duplicate, interpolated from the SPARC standard curve and corrected for sample dilution. Undiluted samples are as follows: serum 10%, plasma (citrate) 20%, plasma (EDTA) 10% and plasma (heparin) 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean SPARC concentration was determined to be 68.59 ng/mL in neat serum, 16.69 ng/mL in neat plasma (citrate), 71.59 ng/mL in neat plasma (EDTA) and 90.38 ng/mL in neat plasma (heparin).
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Interpolated concentrations of native SPARC in human serum samples.Serum from ten individual healthy human male donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean SPARC concentration was determined to be 32.4 ng/mL with a range of 16.78 – 64.84 ng/mL.
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Interpolated concentrations of native SPARC in human plasma samples.Regular and platelet-poor plasma samples were collected per the instructions in the protocol booklet with the matching regular and platelet-poor samples collected at the same time from the same donor. The concentrations of SPARC were measured in duplicates, interpolated from the SPARC standard curves and corrected for sample dilution. Undiluted samples are as follows: plasma (citrate) and platelet-poor plasma (citrate) 20%, plasma (EDTA) and platelet-poor plasma (EDTA) 10%, and plasma (heparin) and platelet-poor plasma heparin 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean SPARC concentration was determined to be 14.79 ng/mL in neat plasma (citrate), 18.95 ng/mL in neat platelet-poor plasma (citrate), 71.56 ng/mL in neat plasma (EDTA), 78.24 ng/mL in neat platelet-poor plasma (EDTA), 119.66 ng/mL in neat plasma (heparin), and 102.81 ng/mL in neat platelet-poor plasma (heparin).
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Sandwich ELISA - Human SPARC ELISA Kit (ab220654)To learn more about the advantages of recombinant antibodies see here.
