Overview

  • Product name

    Rat Clusterin ELISA Kit
    See all Clusterin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Rat plasma 5 5.5%
    Inter-assay
    Sample n Mean SD CV%
    Rat plasma 3 4.2%
  • Sample type

    Serum, Other biological fluids, Hep Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.63 µg/ml
  • Range

    0.8 ng/ml - 50 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 101 96% - 108%
    Other biological fluids 108 101% - 117%
    Hep Plasma 103 99% - 107%
    Cit plasma 110 103% - 123%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Rat
    Does not react with: Cow
  • Product overview

    Clusterin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Clusterin protein in ratserum, plasma, and tissue supernatant samples


    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.


    Clusterin (also known as Apolipoprotein J) is heterodimeric protein involved in a wide variety of cellular functions through its role as a chaperone protein. Clusterin was identified in ram testis fluid and named because of its ability to induce cell clustering. Gene variations in human Clusterin have been associated with Alzheimer's disease, and altered Clusterin levels have been associated with various cancers, hepatitis C, myocardial infarction and other diseases.


     

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Rat Clusterin Capture Antibody 1 x 600µl
    10X Rat Clusterin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BR 1 x 6ml
    Plate Seals 1 unit
    Rat Clusterin Lyophilized Recombinant Protein 2 vials
    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

  • Function

    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.
  • Tissue specificity

    Detected in blood plasma, cerebrospinal fluid, milk, seminal plasma and colon mucosa. Detected in the germinal center of colon lymphoid nodules and in colon parasympathetic ganglia of the Auerbach plexus (at protein level). Ubiquitous. Detected in brain, testis, ovary, liver and pancreas, and at lower levels in kidney, heart, spleen and lung.
  • Sequence similarities

    Belongs to the clusterin family.
  • Post-translational
    modifications

    Isoform 1 is proteolytically cleaved on its way through the secretory system, probably within the Golgi lumen.
    Polyubiquitinated, leading to proteasomal degradation.
    Heavily N-glycosylated. About 30% of the protein mass is comprised of complex N-linked carbohydrate.
  • Cellular localization

    Secreted. Can retrotranslocate from the secretory compartments to the cytosol upon cellular stress and Nucleus. Cytoplasm. Mitochondrion membrane. Cytoplasm, cytosol. Microsome. Endoplasmic reticulum. Cytoplasmic vesicle, secretory vesicle, chromaffin granule. 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.
  • Information by UniProt
  • Alternative names

    • 40
    • AAG 4
    • AAG4
    • Aging associated protein 4
    • Aging-associated gene 4 protein
    • AI893575
    • APO J
    • Apo-J
    • APOJ
    • ApoJalpha
    • ApoJbeta
    • Apolipoprotein J
    • ApolipoproteinJ
    • CLI
    • CLU
    • CLU1
    • CLU2
    • CLUS_HUMAN
    • Clusterin
    • Clusterin alpha chain
    • Clusterin beta chain
    • Complement associated protein SP 40
    • Complement associated protein SP 40 40
    • Complement associated protein SP40
    • Complement cytolysis inhibitor
    • Complement cytolysis inhibitor a chain
    • Complement cytolysis inhibitor b chain
    • Complement lysis inhibitor
    • Complement-associated protein SP-40
    • D14Ucla3
    • Dimeric acid glycoprotein
    • Glycoprotein 80
    • Glycoprotein III
    • GP80
    • Ku70-binding protein 1
    • KUB 1
    • KUB1
    • MGC24903
    • NA1/NA2
    • RATTRPM2B
    • SGP 2
    • SGP2
    • SP 40
    • SP40
    • Sugp-2
    • Sulfated glycoprotein 2
    • Testosterone repressed prostate message 2
    • Testosterone-repressed prostate message 2
    • TRPM 2
    • TRPM-2
    • TRPM2
    • TRPM2B
    • Trpmb
    see all
  • Database links

Associated products

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.

     

  • The Clusterin standard curve was prepared as described in Section 10.

  • The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1: 4,000, plasma (citrate) 1: 4,000, and plasma (heparin) 1: 4,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration was determined to be 87 µg/mL in serum, 102 µg/mL in plasma (citrate), and 119 µg/mL in plasma (heparin).

  • The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: liver supernatant 40% and stomach supernatant 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration was determined to be 102 ng/ml in liver supernatant and 62 ng/mL in stomach supernatant.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration was determined to be 172 µg/mL with a range of 155 – 189 µg/mL.

Protocols

References

ab231934 has not yet been referenced specifically in any publications.

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