Human Clusterin Quantikine ELISA Kit

  (7 citations)     
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Assay Procedure
Citations (7)
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  • 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.
Control Available
QC235 , Quantikine Immunoassay Control Group 235 - Please Inquire
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.
 Clusterin/APOJ [HRP]
Product Datasheets

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Preparation and Storage
  • 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-40,40, 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.
  • 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
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   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.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   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.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.
Citations:

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.

7 Citations: Showing 1 - 7
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Species
Sample Type
  1. 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;0(0):.
    Species: Human
    Sample Type: Cell Culture Supernates
  2. 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;8(0):28-34.
    Species: Human
    Sample Type: Serum
  3. 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;7(1):5035.
    Species: Human
    Sample Type: Plasma
  4. 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, 2016;30(1):124-30.
    Species: Human
    Sample Type: Plasma
  5. 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;55(4):2491-9.
    Species: Human
    Sample Type: Plasma
  6. 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;9(7):e103187.
    Species: Human
    Sample Type: CSF
  7. Semen clusterin is a novel DC-SIGN ligand.
    Authors: Sabatte J, Faigle W, Ceballos A, Morelle W, Rodrígues 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;187(10):5299-309.
    Species: Human
    Sample Type: Seminal Plasma

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