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    products/elisa/mouse-clusterin-elisa-kit-apolipoprotein-j-ab199079.pdf

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Cell Biology Apoptosis Intracellular Associated Proteins
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RecombinantSimpleStep

Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)

  • Datasheet
  • SDS
  • Protocol Booklet
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Other - Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)
  • Example of Clusterin standard curve.
  • Comparison of secreted Clusterin in unstimulated and PMA/PHA-stimulated L929 Cells.
  • Linearity of dilution of the assay.
  • Sandwich ELISA - Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 280 pg/ml
  • Range: 0.78 ng/ml - 50 ng/ml
  • Sample type: Cell culture supernatant, Cit plasma, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Mouse

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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, Cit 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%
    Cit plasma 106 103% - 111%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Sheep, Goat, Cow, Pig
  • Product overview

    Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Clusterin (Apolipoprotein J) protein in cell culture supernatant, cit plasma, serum, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse Clusterin (Apolipoprotein J) with 280 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • 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.

  • 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

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Associated Proteins
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Proteasome
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Other Chaperones
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Signal transduction proteins ELISA kits
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • Ubiquitin Binding
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Nuclear Signaling ELISA kits
    • Neuroscience
    • Diseases
  • 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

    • Entrez Gene: 12759 Mouse
    • SwissProt: Q06890 Mouse

    Associated products

    • Alternative Versions

      • Mouse Clusterin Antibody Pair - BSA and Azide free (ab241706)
    • Related Products

      • Human Clusterin ELISA Kit (ab174447)

    Images

    • Other - Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)
      Other - Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)

      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.

       

    • Example of Clusterin standard curve.
      Example of Clusterin standard curve.

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

    • Comparison of secreted Clusterin in unstimulated and PMA/PHA-stimulated L929 Cells.
      Comparison of secreted Clusterin in unstimulated and PMA/PHA-stimulated L929 Cells.

      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.

    • Linearity of dilution of the assay.
      Linearity of dilution of the assay.

      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.

    • Sandwich ELISA - Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)
      Sandwich ELISA - Mouse Clusterin ELISA Kit (Apolipoprotein J) (ab199079)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab199079? Please let us know so that we can cite the reference in this datasheet.

    ab199079 has been referenced in 2 publications.

    • Xie Z  et al. Monocyte upregulation of podoplanin during early sepsis induces complement inhibitor release to protect liver function. JCI Insight 5:N/A (2020). PubMed: 32641582
    • de Retana SF  et al. Peripheral administration of human recombinant ApoJ/clusterin modulates brain beta-amyloid levels in APP23 mice. Alzheimers Res Ther 11:42 (2019). PubMed: 31077261

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