Human Clusterin ELISA Kit (ab174447)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 22.5 pg/ml
- Range: 234.4 pg/ml - 15000 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human, Rhesus monkey
Overview
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Product name
Human Clusterin ELISA Kit
See all Clusterin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 9 2.7% Inter-assay Sample n Mean SD CV% Serum 3 4.9% -
Sample type
Cell culture supernatant, Saliva, Milk, Urine, Serum, Cell culture extracts, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
22.5 pg/ml -
Range
234.4 pg/ml - 15000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 103 100% - 106% Saliva 100 99% - 100% Milk 99 97% - 101% Urine 102 101% - 102% Serum 101 98% - 103% Cell culture extracts 105 96% - 112% Hep Plasma 102 101% - 103% EDTA Plasma 101 99% - 102% Cit plasma 107 106% - 109% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human, Rhesus monkey
Does not react with: Cow -
Product overview
Human Clusterin ELISA kit (ab174447) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Clusterin protein in human serum, plasma, urine, saliva, milk, cell culture supernatant, and cell extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate human Clusterin with 36.1 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 strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits. -
Notes
Clusterin (CLU) is composed of an antiparallel, disulfide-linked heterodimer of an alpha chain and a beta chain that self-associate and can form higher oligomers. It is ubiquitously expressed in various tissues. Clusterin functions as an extracellular chaperone that prevents aggregation of nonnative proteins. It does not require ATP and does not refold proteins by itself. Clusterin does so by maintaining partially unfolded proteins in a state appropriate for subsequent refolding by other chaperones, such as HSPA8/HSC70. Clusterin prevents stress-induced aggregation of blood plasma proteins by inhibiting the formation of amyloid fibrils through APP, APOC2, B2M, CALCA, CSN3, SNCA and aggregation-prone LYZ variants (in vitro). It can bind to cell surface receptors and trigger internalization of the chaperone-client complex and subsequent lysosomal or proteasomal degradation.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated 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 Clusterin Capture Antibody 1 x 600µl 10X Human Clusterin Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BR 1 x 6ml Human Clusterin 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
Isoform 1 functions as extracellular chaperone that prevents aggregation of nonnative proteins. Prevents stress-induced aggregation of blood plasma proteins. Inhibits formation of amyloid fibrils by APP, APOC2, B2M, CALCA, CSN3, SNCA and aggregation-prone LYZ variants (in vitro). Does not require ATP. Maintains partially unfolded proteins in a state appropriate for subsequent refolding by other chaperones, such as HSPA8/HSC70. Does not refold proteins by itself. Binding to cell surface receptors triggers internalization of the chaperone-client complex and subsequent lysosomal or proteasomal degradation. Secreted isoform 1 protects cells against apoptosis and against cytolysis by complement. Intracellular isoforms interact with ubiquitin and SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complexes and promote the ubiquitination and subsequent proteasomal degradation of target proteins. Promotes proteasomal degradation of COMMD1 and IKBKB. Modulates NF-kappa-B transcriptional activity. Nuclear isoforms promote apoptosis. Mitochondrial isoforms suppress BAX-dependent release of cytochrome c into the cytoplasm and inhibit apoptosis. Plays a role in the regulation of cell proliferation. -
Tissue specificity
Detected in blood plasma, cerebrospinal fluid, milk, seminal plasma and colon mucosa. Detected in the germinal center of colon lymphoid nodules and in colon parasympathetic ganglia of the Auerbach plexus (at protein level). Ubiquitous. Detected in brain, testis, ovary, liver and pancreas, and at lower levels in kidney, heart, spleen and lung. -
Sequence similarities
Belongs to the clusterin family. -
Post-translational
modificationsIsoform 1 is proteolytically cleaved on its way through the secretory system, probably within the Golgi lumen.
Polyubiquitinated, leading to proteasomal degradation.
Heavily N-glycosylated. About 30% of the protein mass is comprised of complex N-linked carbohydrate. -
Cellular localization
Secreted. Can retrotranslocate from the secretory compartments to the cytosol upon cellular stress and Nucleus. Cytoplasm. Mitochondrion membrane. Cytoplasm, cytosol. Microsome. Endoplasmic reticulum. Cytoplasmic vesicle, secretory vesicle, chromaffin granule. Isoforms lacking the N-terminal signal sequence have been shown to be cytoplasmic and/or nuclear. Secreted isoforms can retrotranslocate from the secretory compartments to the cytosol upon cellular stress. Detected in perinuclear foci that may be aggresomes containing misfolded, ubiquitinated proteins. Detected at the mitochondrion membrane upon induction of apoptosis. - Information by UniProt
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Alternative names
- 40
- AAG 4
- AAG4
see all -
Database links
- Entrez Gene: 1191 Human
- Omim: 185430 Human
- SwissProt: P10909 Human
- Unigene: 436657 Human
Images
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Other - Human Clusterin ELISA Kit (ab174447)
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 Clusterin standard curve in Sample Diluent NS.The Clusterin standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Example of human Clusterin standard curve in 1X Cell Extraction Buffer PTR.The Clusterin standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Clusterin in human serum and plasma samples.The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:80,000, plasma (heparin) 1:40,000, plasma (EDTA) 1:40,000, and plasma (citrate) 1:20,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration in the neat samples was determined to be 174 µg/mL in serum, 123 µg/mL in plasma (heparin), 158 µg/mL in plasma (EDTA), and 206 µg/mL in plasma (citrate).
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Interpolated concentrations of native Clusterin in human urine, saliva, and milk.The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 1:32, saliva 1:80, and milk 1:3,200. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration in the neat samples was determined to be 157 ng/mL in urine, 246 ng/mL in saliva, and 20,323 ng/mL in milk.
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Interpolated concentrations of native Clusterin in human HepG2 cell supernatant, HepG2 cell extract, and MDA-MB-231 cell extract.The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: HepG2 supernatant 1:12, HepG2 extract 200 µg/mL, and MDA-MB-231 extract 750 µg/mL.
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Interpolated concentrations of native Clusterin in Rhesus Macaque plasma (EDTA).The concentrations of Clusterin were measured in duplicate, interpolated from the Clusterin standard curve and corrected for sample dilution. Undiluted samples are 1:20,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration in the neat sample was determined to be 94 µg/mL.
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Serum from ten individual healthy human female donors was measured in duplicate.Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration was determined to be 150 µg/mL with a range of 92 – 227 µg/mL.
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Human Clusterin Competitor Std Curve ComparisonStandard curve comparison between human Clusterin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.
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Kit Comparison between current and original versionStandard curve comparison between current human Clutsterin SimpleStep ELISA® kit and original human Clusterin SimpleStep ELISA® kit. The current SimpleStep ELISA kit shows increased sensitivity.
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Sandwich ELISA - Human Clusterin ELISA Kit (ab174447)To learn more about the advantages of recombinant antibodies see here.