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Immunology Innate Immunity Complement Classical Pathway
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)

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  • Certificate of Compliance
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Western blot - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
  • Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2911] to Clusterin - Low endotoxin, Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

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Conjugation
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HRP Conjugation Kit - Lightning-Link® (ab102890)
Protein
Recombinant Mouse Clusterin protein (ab194035)

View more associated products

Overview

  • Product name

    Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free
    See all Clusterin primary antibodies
  • Description

    Rabbit monoclonal [EPR2911] to Clusterin - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    aa 400-500.

  • Positive control

    • IHC-P: human breast carcinoma and testis tissue; WB: human testis, brain, tonsil, fetal brain and plasma lysates
  • General notes

    ab229445 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2911
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Complement
    • Classical Pathway
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipoproteins/Apolipoproteins
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cancer
    • Tumor biomarkers
    • Other
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipoproteins/Apolipoproteins
    • Apolipoproteins

Associated products

  • Alternative Versions

    • Anti-Clusterin antibody [EPR2911] (ab92548)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Mouse Clusterin protein (ab194035)

Applications

Our Abpromise guarantee covers the use of ab229445 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

WB Use at an assay dependent concentration. Predicted molecular weight: 52 kDa.

Target

  • 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.
  • Target information above from: UniProt accession P10909 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1191 Human
    • Omim: 185430 Human
    • SwissProt: P10909 Human
    • Unigene: 436657 Human
    • Alternative names

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

    Images

    • Western blot - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
      Western blot - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
      All lanes : Anti-Clusterin antibody [EPR2911] (ab92548) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : Clusterin knockout HAP1 whole cell lysate
      Lane 3 : U87-MG whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 52 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab92548 observed at 68 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab92548 was shown to recognize Clusterin in wild-type HAP1 cells as signal was lost at the expected MW in Clusterin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Clusterin knockout samples were subjected to SDS-PAGE. Ab92548 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229445).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Clusterin with purified ab92548 at 1/450 dilution (0.23 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92548)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)

      This IHC data was generated using the same anti-Clusterin antibody clone, EPR2911, in a different buffer formulation (cat# ab925480).

      ab92548 (unpurified) at 1/100 dilution staining Apolipoprotein J in Human testis tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.

      Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

    • Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
      Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab229445? Please let us know so that we can cite the reference in this datasheet.

    ab229445 has not yet been referenced specifically in any publications.

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