Mouse Clusterin Quantikine ELISA Kit

  (2 citations)     
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Product Details
Assay Procedure
Citations (2)
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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), Cell Lysates (10 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (10 uL)
  • Sensitivity
    0.094 ng/mL
  • Assay Range
    0.8 - 50 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant mouse 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.
Product Summary
The Quantikine Mouse Clusterin Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse Clusterin levels in cell culture supernates, cell lysates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse Clusterin and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse Clusterin showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse 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, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.35 8.02 17.5 1.36 7.62 16.2
Standard Deviation 0.086 0.282 0.682 0.119 0.577 1.62
CV% 6.4 3.5 3.9 8.8 7.6 10

Recovery

The recovery of mouse Clusterin spiked into various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 101 90-111
Cell Lysates (n=4) 106 95-117
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse Clusterin were serially diluted with 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. 50 µL Assay Diluent
  4.   Add 50 µL of the appropriate 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 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 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 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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.

2 Citations: Showing 1 - 2
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Species
Sample Type
  1. Clusterin Seals the Ocular Surface Barrier in Mouse Dry Eye.
    Authors: Bauskar A, Mack W, Mauris J, Argueso P, Heur M, Nagel B, Kolar G, Gleave M, Nakamura T, Kinoshita S, Moradian-Oldak J, Panjwani N, Pflugfelder S, Wilson M, Fini M, Jeong S
    PLoS ONE, 2015;10(9):e0138958.
    Species: Mouse
    Sample Type: Tears
  2. Analysis of the hippocampal proteome in ME7 prion disease reveals a predominant astrocytic signature and highlights the brain-restricted production of clusterin in chronic neurodegeneration.
    Authors: Asuni A, Gray B, Bailey J, Skipp P, Perry V, O'Connor V
    J Biol Chem, 2014;289(7):4532-45.
    Species: Mouse
    Sample Type: Plasma

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