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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (25 uL), Urine (13 uL)
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Sensitivity
1.05 ng/mL
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Assay Range
3.1 - 200 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
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Specificity
Natural and recombinant human Clusterin
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC235, Quantikine Immunoassay Control Group 235 -
Product Summary
The Quantikine Human Clusterin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Clusterin in cell culture supernates, serum, plasma, urine, and saliva. It contains NS0-expressed recombinant human Clusterin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Clusterin 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 Clusterin.
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, EDTA Plasma, Heparin Plasma, Saliva, Urine | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 19 | 64.3 | 130 | 22.1 | 64.1 | 136 |
| Standard Deviation | 0.7 | 2.4 | 4.5 | 1.6 | 5.4 | 9.3 |
| CV% | 3.7 | 3.7 | 3.5 | 7.2 | 8.4 | 6.8 |
Recovery
The recovery of Clusterin spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=4) | 107 | 102-110 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of Clusterin were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: Clusterin
Clusterin, also known as Apolipoprotein J, Sulfated Glycoprotein 2 (SGP-2), TRPM-2, and SP-40,40, is a secreted multifunctional protein that binds a wide range of molecules and may function as a chaperone of misfolded extracellular proteins. It also participates in the control of cell proliferation, apoptosis, and carcinogenesis. Clusterin is predominantly expressed in adult testis, ovary, adrenal gland, liver, heart, and brain and in many epithelial tissues during embryonic development. It is secreted as a disulfide-linked heterodimer of alpha and beta chains and circulates as a component of high density lipoprotein (HDL) particles. Clusterin exerts tissue protective functions in atherosclerosis and Alzheimer’s disease by binding and neutralizing non-oxidatively modified LDL reduces cytotoxicity and beta-amyloid fibrils, respectively. Increased circulating levels of Clusterin enhance tumor aggressiveness by inhibiting apoptosis and by promoting epithelial to mesenchymal transition.
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Alternate Names
40; 40, sulfated glycoprotein 2; Aging-associated gene 4 protein; aging-associated protein 4; APOJ; Apo-J; Apolipoprotein J; CLI; CLIclusterin (complement lysis inhibitor, SP-40; CLU; Clusterin; Complement cytolysis inhibitor; complement lysis inhibitor; Complement-associated protein SP-40; Ku70-binding protein 1; KUB1SGP2; MGC24903; NA1/NA2; SGP-2; SP-40; sulfated glycoprotein 2; Testosterone-repressed prostate message 2; testosterone-repressed prostate message 2, apolipoprotein J); TRPM-2; TRPM-2TRPM2;
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