Overview

  • Product name

    Mouse Clusterin ELISA Kit (Apolipoprotein J)
    See all Apolipoprotein J (Clusterin) kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 8.2%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 12.8%
  • Sample type

    Cell culture supernatant, Urine, Serum, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    280 pg/ml
  • Range

    0.78 ng/ml - 50 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 102 95% - 105%
    Serum 105 97% - 112%
    Cell culture media 99 95% - 110%
    Citrate Plasma 106 103% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Sheep, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Human, Pig
  • Product overview

    Clusterin (Apolipoprotein J) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Clusterin protein in mouse serum, plasma, urine and cell culture supernatant.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Clusterin (also known as Apoliprotein J) is a multifunctional protein, named for its ability to induce cellular clustering. The secreted form is processed from a single chain into alpha and beta strands, that form a heterodimeric, 80 kDa protein. Clusterin is also present as an alternatively spliced, 50 kDa intracellular form isoform. As an extracellular chaperone, Clusterin prevents the aggregation of blood plasma proteins and inhibits amyloid fibrils. Clusterin also plays a role in both apoptosis and proliferation. Secreted Clusterin protects against apoptosis by the complement system, but can promote apoptosis when present in the nucleus. Intracellular Clusterin interacts with ubiquitin and the proteasomal machinery for target degradation.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Clusterin Capture Antibody 1 x 600µl
    10X Mouse Clusterin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Mouse 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

  • Relevance

    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.
  • Cellular localization

    Isoform 1: Secreted. Note: Can retrotranslocate from the secretory compartments to the cytosol upon cellular stress. Nucleus. Cytoplasm. Mitochondrion membrane; Peripheral membrane protein; Cytoplasmic side. Cytoplasm › cytosol. Microsome. Endoplasmic reticulum. Cytoplasmic vesicle › secretory vesicle › chromaffin granule. Note: 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.
  • Alternative names

    • AAG4
    • Aging associated protein 4
    • APO J
    • APOJ
    • CLI
    • CLU
    • CLU1
    • CLU2
    • Clusterin
    • Complement associated protein SP 40,40
    • Complement cytolysis inhibitor
    • Complement lysis inhibitor
    • Ku70 binding protein 1
    • KUB1
    • NA1/NA2
    • SGP 2
    • SGP2
    • SP 40
    • SP40
    • Sulfated glycoprotein 2
    • Testosterone repressed prostate message 2
    • TRPM 2
    • TRPM2
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab199079 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • L929 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. Clusterin was measured in 2-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated L929 and cell culture media. Measured values were interpolated from the Clusterin Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 1.5 ng/mL unstimulated, 16.4 ng/mL stimulated. There was no detectable signal in media.

  • Interpolated concentrations of Clusterin in mouse serum and plasma (citrate). The concentrations of Clusterin were measured in duplicate and interpolated from the Clusterin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration was determined to be 59.1 µg/mL in mouse serum and 38.7 µg /mL in mouse plasma.

Protocols

References

ab199079 has not yet been referenced specifically in any publications.

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