Summary
Sample Values
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human Clusterin Quantikine ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
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)
Sensitivity
1.05 ng/mL
Assay Range
3.1 - 200 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human Clusterin
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for the presence of human Clusterin in this assay. No medical histories were available for the donors used in this study.| Sample Type | Mean (μg/mL) | Range (μg/mL) | Standard Deviation (μg/mL) |
| Serum (n=36) | 266 | 195-378 | 42.8 |
| Heparin plasma (n=36) | 266 | 190-377 | 44.6 |
| EDTA plasma (n=36) | 261 | 185-436 | 52.0 |
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Salive (n=17) | 334 | 85.2-624 | 176 |
| Urine (n=21) | 280 | 85.2-628 | 171 |
Cell Culture Supernates:
Human peripheral blood leukocytes (PBLs) were cultured in DMEM supplemented with
5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL
PHA. Aliquots of the cell culture supernates were removed and assayed for levels of human
Clusterin. All samples were non-detectable.
HepG2 human hepatocellular carcinoma cells were cultured in MEM (NEAA salts)
supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine,
100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture
supernate was removed, assayed for human Clusterin, and measured 574 ng/mL.
HeLa cervical epithelial carcinoma cells were cultured in RPMI supplemented with 10% fetal
bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed
for human Clusterin, and measured 863 ng/mL.
HUVEC human umbilical vein endothelial cells were cultured in EGM-2 media and stimulated
with recombinant human IL-1 beta for 24 hours. An aliquot of the cell culture supernate was
removed, assayed for human Clusterin, and measured 17.3 ng/mL.
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 (pg/mL) | 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.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Clusterin
Clusterin [also known as Apolipoprotein J, Sulfated Glycoprotein 2 (SGP-2), TRPM-2, and SP-40], is a secreted multifunctional protein that was named for its ability to induce cellular clustering. It 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 (1, 2). Clusterin is predominantly expressed in adult testis, ovary, adrenal gland, liver, heart, brain, and in many epithelial tissues during embryonic development (3). Human Clusterin is synthesized as a precursor that contains two coiled coil domains, three nuclear localization signals (NLS), and one heparin binding domain (4 - 6). Intracellular cleavages of the precursor remove the signal peptide and generate comparably sized alpha and beta chains which are secreted as an approximately 80 kDa N-glycosylated and disulfide-linked heterodimer (7-9). Mature human Clusterin shares a 77% amino acid sequence identity with mouse and rat Clusterin.
High μg/mL concentrations of Clusterin circulate predominantly as a component of high
density lipoprotein particles, and these are internalized and degraded through interactions
with LRP-2/Megalin (10, 11). The ability of Clusterin to bind and neutralize non-oxidatively
modified LDL reduces cytotoxicity in atherosclerotic plaques (12). The chaperone function of
Clusterin helps to reduce the accumulation of beta -amyloid fibrils and damage due to amyloid
plaques in Alzheimer's disease (13). An alternately spliced 50 kDa isoform of human Clusterin
(nCLU) remains intracellular and is neither glycosylated nor cleaved into alpha and beta chains (5, 14).
Cellular exposure to ionizing radiation promotes the translocation of nCLU to the nucleus
where it interacts with Ku70 and promotes apoptosis (5, 14). This function contrasts with the
cytoprotective effect of secreted Clusterin (15). During tumor progression, nCLU is down
regulated while the secreted form is upregulated and may be aberrantly glycosylated (14, 16,
17). Increased circulating levels of Clusterin enhance tumor aggressiveness by inhibiting
apoptosis and by promoting the epithelial to mesenchymal transition (18-20).
Entrez Gene IDs:
1191 (Human); 12759 (Mouse)
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Human Clusterin Quantikine ELISA Kit
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Alzheimer's disease protective allele of Clusterin modulates neuronal excitability through lipid-droplet-mediated neuron-glia communication
Authors: Zhao, X;Li, Y;Zhang, S;Sudwarts, A;Zhang, H;Kozlova, A;Moulton, MJ;Goodman, LD;Pang, ZP;Sanders, AR;Bellen, HJ;Thinakaran, G;Duan, J;
medRxiv : the preprint server for health sciences
Species: Human
Sample Types: Cell Culture Supernates
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Utility of Three Serum Biomarkers for Early Detection of Systemic Envenoming Following Viper Bites in Sri Lanka
Authors: Wedasingha, S;Silva, A;Fakes, K;Siribaddana, S;Isbister, GK;
Annals of emergency medicine
Species: Human
Sample Types: Serum
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Serum and urinary biomarkers of vancomycin-induced acute kidney injury: A prospective, observational analysis
Authors: Park, SI;Yu, U;Oh, WS;Ryu, SW;Son, S;Lee, S;Baek, H;Park, JI;
Medicine
Species: Human
Sample Types: Urine
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Clinical usefulness of urinary biomarkers for early prediction of acute kidney injury in patients undergoing transaortic valve implantation
Authors: Obata, Y;Kamijo-Ikemori, A;Shimmi, S;Inoue, S;
Scientific reports
Species: Human
Sample Types: Urine
-
Metabolic Syndrome, Clusterin and Elafin in Patients with Psoriasis Vulgaris
Authors: D Holmannova, P Borsky, L Borska, C Andrys, K Hamakova, V Rehacek, T Svadlakova, A Malkova, M Beranek, V Palicka, J Krejsek, Z Fiala
Int J Mol Sci, 2020-08-05;21(16):.
Species: Human
Sample Types: Serum
-
Clusterin from human clinical tear samples: Positive correlation between tear concentration and Schirmer strip test results
Authors: V Yu, D Bhattachar, A Webster, A Bauskar, C Flowers, M Heur, SK Chintala, T Itakura, MR Wilson, JT Barr, S Jeong, M Wang, ME Fini
Ocul Surf, 2018-08-02;0(0):.
Species: Human
Sample Types: Protein
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Relevance of N-terminal residues for amyloid-? binding to platelet integrin ?IIb?3, integrin outside-in signaling and amyloid-? fibril formation
Authors: L Donner, L Gremer, T Ziehm, CGW Gertzen, H Gohlke, D Willbold, M Elvers
Cell. Signal., 2018-06-30;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Role of clusterin/progranulin in toluene diisocyanate-induced occupational asthma
Authors: GS Choi, HKT Trinh, EM Yang, YM Ye, YS Shin, SH Kim, HS Park
Exp. Mol. Med., 2018-05-01;50(5):58.
Species: Human
Sample Types: Serum
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Plasma biomarkers are associated with agitation and regional brain atrophy in Alzheimer's disease
Authors: JL Hsu, WJ Lee, YC Liao, JF Lirng, SJ Wang, JL Fuh
Sci Rep, 2017-07-11;7(1):5035.
Species: Human
Sample Types: Plasma
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Novel panel of protein biomarkers to predict response to bortezomib-containing induction regimens in multiple myeloma patients
Authors: KR Ting, M Henry, J Meiller, A Larkin, M Clynes, P Meleady, D Bazou, P Dowling, P O'Gorman
BBA Clin, 2017-06-07;8(0):28-34.
Species: Human
Sample Types: Serum
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Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-Related Macular Degeneration
Authors: JS Saini, B Corneo, JD Miller, TR Kiehl, Q Wang, NC Boles, TA Blenkinsop, JH Stern, S Temple
Cell Stem Cell, 2017-01-26;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Increased IL6 plasma levels in indolent systemic mastocytosis patients are associated with high risk of disease progression.
Authors: Mayado A, Teodosio C, Garcia-Montero A, Matito A, Rodriguez-Caballero A, Morgado J, Muniz C, Jara-Acevedo M, Alvarez-Twose I, Sanchez-Munoz L, Matarraz S, Caldas C, Munoz-Gonzalez J, Escribano L, Orfao A
Leukemia, 2015-07-08;30(1):124-30.
Species: Human
Sample Types: Plasma
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Intracellular clusterin interacts with brain isoforms of the bridging integrator 1 and with the microtubule-associated protein Tau in Alzheimer's disease.
Authors: Zhou Y, Hayashi I, Wong J, Tugusheva K, Renger J, Zerbinatti C
PLoS ONE, 2014-07-22;9(7):e103187.
Species: Human
Sample Types: CSF
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Clusterin and complement activation in exfoliation glaucoma.
Authors: Doudevski, Ivo, Rostagno, Agueda, Cowman, Mary, Liebmann, Jeffrey, Ritch, Robert, Ghiso, Jorge
Invest Ophthalmol Vis Sci, 2014-04-17;55(4):2491-9.
Species: Human
Sample Types: Plasma
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Semen clusterin is a novel DC-SIGN ligand.
Authors: Sabatte J, Faigle W, Ceballos A, Morelle W, RodrÃgues CR, Lenicov FR, Thepaut M, Fieschi F, Malchiodi E, Fernandez M, Arenzana-Seisdedos F, Lortat-Jacob H, Michalski JC, Geffner J, Amigorena S
J. Immunol., 2011-10-17;187(10):5299-309.
Species: Human
Sample Types: Seminal Plasma
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Human Clusterin Quantikine ELISA Kit
By Anonymous
on 04/21/2022
Sample Tested: Urine
Human Clusterin Quantikine ELISA Kit
By Anonymous
on 11/10/2020
Sample Tested: putification steps
Human Clusterin Quantikine ELISA Kit
By Anonymous
on 11/27/2018
Sample Tested: Serum and Plasma
