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Catalog No: OKCD01150 (Formerly GWB-KBBEZ8)
Size:96WELLS
Price: $650.00
SKU
OKCD01150
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Datasheets/ManualsPrintable datasheet for SPARC ELISA Kit (Rat) (OKCD01150)
Product Info
Predicted Species ReactivityRat|Rattus norvegicus
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit Linearity
Sample1:2 Dilution1:4 Dilution1:8 Dilution1:16 Dilution
serum(n=5)81-91%87-101%94-105%93-105%
EDTA plasma(n=5)80-101%82-92%89-97%85-105%
heparin plasma(n=5)97-105%91-105%98-105%93-105%
ELISA Kit PrincipleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Osteonectin (ON). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Osteonectin (ON). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Osteonectin (ON), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Osteonectin (ON) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range6.25-400ng/mL
ELISA Kit Recovery
MatrixRecovery Range (%)Average (%)
serum(n=5)79-9083
EDTA plasma(n=5)78-10591
heparin plasma(n=5)86-9591
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteonectin (ON) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Osteonectin (ON) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
Anti-Sparc Microplate96 Wells (12 x 8 Well Strips)
Sparc Lyophilized Standard2 x 40 ng
100X Biotinylated Sparc Detector Antibody120 uL
Avidin/HRP Conjugate120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
TMB Substrate1 x 9 mL
Stop Solution1 x 6 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Sensitivity< 0.242 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of Osteonectin (ON). No significant cross-reactivity or interference between Osteonectin (ON) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich Immunoassay
Gene SymbolSparc
Gene Full Namesecreted protein acidic and cysteine rich
Alias Symbolsbasement-membrane protein 40;BM-40;ON;osteonectin;secreted acidic cysteine rich glycoprotein;Secreted acidic cystein-rich glycoprotein (osteonectin);secreted protein acidic and rich in cysteine;secreted protein, acidic, cysteine-rich (osteonectin);SPARC.
NCBI Gene Id24791
Protein NameSPARC
Description of TargetAppears 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.
Uniprot IDP16975
Protein Accession #NP_033268
Nucleotide Accession #NM_009242.5
Protein Size (# AA)301
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