Human SPARC ELISA Kit (ab220654)
- Datasheet
- References
- Protocols
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
SPARC in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of human SPARC protein in serum, plasma, and cell culture supernatant samples.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
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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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Tested applications
Suitable for: Sandwich ELISAmore details -
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 SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 x 96 tests Sample Diluent NS (ab193972) 1 x 50ml 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
Applications
Our Abpromise guarantee covers the use of ab220654 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
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).
Protocols
References
ab220654 has not yet been referenced specifically in any publications.
