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Mouse Clusterin ELISA Kit - Quantikine

Catalog #: MCLU00
4 Citations Datasheet / COA / SDS
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MCLU00
Mouse Clusterin ELISA Standard Curve
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Product Details
Procedure
Citations (4)
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Mouse 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), 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 (pg/mL) 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.
Mouse Clusterin ELISA Linearity

Scientific Data

N/A Mouse Clusterin ELISA Standard Curve View Larger

Mouse Clusterin ELISA Standard Curve

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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 expressed in adult testis, ovary, adrenal gland, liver, heart, brain, and in many epithelial tissues during embryonic development (3). Mouse Clusterin is synthesized as a precursor that contains two coiled coil domains, two 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 mouse Clusterin shares 77% and 93% amino acid sequence identity with human and rat Clusterin, respectively. 

An alternately spliced 50 kDa isoform of mouse Clusterin (nCLU) remains intracellular and is neither glycosylated nor cleaved into alpha and beta chains (4). Cellular exposure to ionizing radiation promotes the translocation of nCLU to the nucleus where it interacts with Ku70 and promotes apoptosis (4, 10). This function contrasts with the cytoprotective effect of secreted Clusterin (11). 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 (12, 13). The ability of Clusterin to bind and neutralize non-oxidatively modified LDL reduces cytotoxicity in atherosclerotic plaques (14). The chaperone function of Clusterin also helps to reduce the accumulation of beta -amyloid fibrils and damage due to amyloid plaques in Alzheimer's disease (15). Clusterin levels are elevated in the cerebrospinal fluid of patients with Alzheimer's disease, Parkinson's disease, and multiple sclerosis (9, 16, 17) and in the urine of patients with kidney injury or bladder cancer (18-20). Clusterin is released by activated platelets at sites of vascular injury and is found in the synovial fluid of rheumatoid arthritis and osteoarthritis patients (21, 22). During human tumor progression, nCLU is downregulated while the secreted form is upregulated and may be aberrantly glycosylated (10, 23-25). Increased circulating levels of Clusterin enhance tumor aggressiveness by inhibiting apoptosis and by promoting epithelial to mesenchymal transition (26-28).

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
&#9888; 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 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
  3.   Add 50 µL of the appropriate Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   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.
  7.   Aspirate and wash 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   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 for Mouse 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.

4 Citations: Showing 1 - 4
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  1. Apolipoprotein�J is a hepatokine regulating muscle glucose metabolism and insulin sensitivity
    Authors: JA Seo, MC Kang, WM Yang, WM Hwang, SS Kim, SH Hong, JI Heo, A Vijyakumar, L Pereira de, A Uner, H Huang, SH Lee, IS Lima, KS Park, MS Kim, Y Dagon, TE Willnow, V Aroda, TP Ciaraldi, RR Henry, YB Kim
    Nat Commun, 2020-04-24;11(1):2024.
    Species: Mouse
    Sample Types: Serum
  2. Concomitant memantine and Lactobacillus plantarum treatment attenuates cognitive impairments in APP/PS1 mice
    Authors: QJ Wang, YE Shen, X Wang, S Fu, X Zhang, YN Zhang, RT Wang
    Aging (Albany NY), 2020-01-06;12(1):628-649.
    Species: Mouse
    Sample Types: Plasma
  3. 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, 2015-09-24;10(9):e0138958.
    Species: Mouse
    Sample Types: Tears
  4. 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, 2013-12-23;289(7):4532-45.
    Species: Mouse
    Sample Types: Plasma

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