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    clusterin-antibody-epr2911-ab92548.pdf

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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] (ab92548)

  • Datasheet
  • SDS
Reviews (3) Submit a question References (6)

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Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] (ab92548)
  • Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] (ab92548)
  • Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
  • Anti-Clusterin antibody [EPR2911] (ab92548)

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
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Protein
Recombinant Mouse Clusterin protein (ab194035)

View more associated products

Overview

  • Product name

    Anti-Clusterin antibody [EPR2911]
    See all Clusterin primary antibodies
  • Description

    Rabbit monoclonal [EPR2911] to Clusterin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • 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
    • Metabolism
    • Types of disease
    • Cancer
    • Neuroscience
    • Diseases

Associated products

  • Alternative Versions

    • Anti-Clusterin antibody [EPR2911] - Low endotoxin, Azide free (ab229445)
    • Anti-Clusterin antibody [EPR2911] - BSA and Azide free (ab271867)
  • Isotype control

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

    • Recombinant Mouse Clusterin protein (ab194035)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab92548 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
WB (1)
1/1000 - 1/2000. Predicted molecular weight: 52 kDa.
IHC-P
1/100 - 1/450. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Notes
WB
1/1000 - 1/2000. Predicted molecular weight: 52 kDa.
IHC-P
1/100 - 1/450. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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] (ab92548)
      Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: Clusterin knockout HAP1 whole cell lysate (20 µg)
      Lane 3: U87-MG whole cell lysate (20 µg)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] (ab92548)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] (ab92548)

      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.

    • Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
      Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
      All lanes : Anti-Clusterin antibody [EPR2911] (ab92548) at 1/1000 dilution (Purified)

      Lane 1 : Human testis lysates
      Lane 2 : Human brain lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 52 kDa
      Observed band size: 37,68 kDa why is the actual band size different from the predicted?



      Clusterin precursor could be cleaved to produce an N-terminal α-chain and a C-terminal β-chain as was described by PMID: 25402950. ab92548 recognizes α-chain.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] (ab92548)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clusterin antibody [EPR2911] (ab92548)

      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.

    • Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
      Western blot - Anti-Clusterin antibody [EPR2911] (ab92548)
      All lanes : Anti-Clusterin antibody [EPR2911] (ab92548) at 1/1000 dilution (unpurified)

      Lane 1 : Human tonsil tissue lysate
      Lane 2 : Fetal brain tissue lysate
      Lane 3 : Human Plasma lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 52 kDa

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

    Protocols

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (6)

    Publishing research using ab92548? Please let us know so that we can cite the reference in this datasheet.

    ab92548 has been referenced in 6 publications.

    • Huang Y  et al. Reduced Expression of Antimicrobial Protein Secretory Leukoprotease Inhibitor and Clusterin in Chronic Rhinosinusitis with Nasal Polyps. J Immunol Res 2021:1057186 (2021). PubMed: 33506054
    • Yuste-Checa P  et al. The extracellular chaperone Clusterin enhances Tau aggregate seeding in a cellular model. Nat Commun 12:4863 (2021). PubMed: 34381050
    • Praktiknjo SD  et al. Tracing tumorigenesis in a solid tumor model at single-cell resolution. Nat Commun 11:991 (2020). PubMed: 32080185
    • Bannai T  et al. Chronic cerebral hypoperfusion shifts the equilibrium of amyloid ß oligomers to aggregation-prone species with higher molecular weight. Sci Rep 9:2827 (2019). PubMed: 30808940
    • Keyvani K  et al. Higher levels of kallikrein-8 in female brain may increase the risk for Alzheimer's disease. Brain Pathol 28:947-964 (2018). PubMed: 29505099
    • Walters MS  et al. Smoking accelerates aging of the small airway epithelium. Respir Res 15:94 (2014). IHC-P ; Human . PubMed: 25248511

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Clusterin antibody [EPR2911]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Primary astrocyte culture)
    Permeabilization
    Yes - 0.1M PBS with 1% Triton X
    Specification
    Primary astrocyte culture
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Methanol
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Sep 06 2016

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Clusterin antibody [EPR2911]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.1 M PBS with 1% Triton X
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Feb 29 2016

    Western blot abreview for Anti-Clusterin antibody [EPR2911]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain lysate)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    30 µg
    Specification
    Brain lysate
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Feb 05 2016

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