Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Calnexin antibody [EPR3632] (ab92573)

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Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] (ab92573)
  • Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3632] (ab92573)
  • Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
  • Anti-Calnexin antibody [EPR3632] (ab92573)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3632] to Calnexin
  • Suitable for: ICC/IF, WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Calnexin antibody [EPR3632]
    See all Calnexin primary antibodies

  • Description

    Rabbit monoclonal [EPR3632] to Calnexin

  • Host species

    Rabbit

  • Specificity

    Recognizes ER membrane, mitochondria and cis-Golgi

  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Calnexin aa 1-100. The exact sequence is proprietary.
    Database link: P27824

  • Positive control

    • WB: HeLa, A431, SH-SY5Y and HepG2 whole cell lysate (ab7900). IHC-P: Human tonsil tissue. ICC/IF: Wild-type HAP1 cells.

  • General notes

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

    References regarding specificity:

    Horner SM  et al. Mitochondrial-associated endoplasmic reticulum membranes (MAM) form innate immune synapses and are targeted by hepatitis C virus. Proc Natl Acad Sci U S A 108:14590-5 (2011). PubMed: 21844353

    Myhill N  et al. The subcellular distribution of calnexin is mediated by PACS-2. Mol Biol Cell 19:2777-88 (2008). PubMed: 18417615

    Yoshimura SI  et al. Direct targeting of cis-Golgi matrix proteins to the Golgi apparatus. J Cell Sci 114:4105-15 (2001). PubMed: 11739642

    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.

    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. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.

  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    EPR3632

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab92573 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
ICC/IF 1/1000.
WB 1/20000 - 1/100000. Predicted molecular weight: 90 kDa.
IP 1/50.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins.

    • Sequence similarities

      Belongs to the calreticulin family.

    • Cellular localization

      Endoplasmic reticulum membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
    • Information by UniProt

    • Database links

      • Alternative names

        • Calnexin antibody
        • CALX_HUMAN antibody
        • CANX antibody
        • CNX antibody
        • FLJ26570 antibody
        • Histocompatibility complex class I antigen binding protein p88 antibody
        • IP90 antibody
        • Major histocompatibility complex class I antigen-binding protein p88 antibody
        • p90 antibody
        see all

      Images

      • Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
        Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
        All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution

        Lane 1 : Wild-type HAP1 cell lysate
        Lane 2 : Calnexin knockout HAP1 cell lysate
        Lane 3 : THP-1 cell lysate
        Lane 4 : RAW 264.7 cell lysate

        Lysates/proteins at 20 µg per lane.

        Predicted band size: 90 kDa



        Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa.

        ab92573 was shown to specifically react with Calnexin when Calnexin knockout samples were used. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab92573 and ab8245 (loading control to GAPDH) were diluted at 1/20,000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

      • Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] (ab92573)
        Immunocytochemistry/ Immunofluorescence - Anti-Calnexin antibody [EPR3632] (ab92573)

        ab92573 staining Calnexin in wild-type HAP1 cells (top panel) and CANX knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92573 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

        Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      • Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
        Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
        All lanes : Anti-Calnexin antibody [EPR3632] (ab92573) at 1/20000 dilution

        Lane 1 : HeLa cell lysate
        Lane 2 : A431 cell lysate
        Lane 3 : SH-SY5Y cell lysate
        Lane 4 : HepG2 cell lysates

        Lysates/proteins at 10 µg per lane.

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

        Predicted band size: 90 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3632] (ab92573)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calnexin antibody [EPR3632] (ab92573)

        Immunohistochemical analysis of paraffin embedded Human tonsil tissue using ab92573 at a 1/100 dilution.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
        Western blot - Anti-Calnexin antibody [EPR3632] (ab92573)
        All lanes : Anti-Calnexin antibody [EPR3632] (ab92573)

        Lane 1 : Wild-type HAP1 cell lysate
        Lane 2 : Calnexin knockout HAP1 cell lysate

        Lysates/proteins at 20 µg per lane.

        Predicted band size: 90 kDa



        Lanes 1 - 2: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa.

        This western blot image is a comparison between ab92573 and a competitor's top cited rabbit polyclonal antibody.

      • Anti-Calnexin antibody [EPR3632] (ab92573)
        Anti-Calnexin antibody [EPR3632] (ab92573)

      References (9)

      Publishing research using ab92573? Please let us know so that we can cite the reference in this datasheet.

      ab92573 has been referenced in 9 publications.

      • Xie M  et al. Exosomal circSHKBP1 promotes gastric cancer progression via regulating the miR-582-3p/HUR/VEGF axis and suppressing HSP90 degradation. Mol Cancer 19:112 (2020). PubMed: 32600329
      • Yan L & Wu X Exosomes produced from 3D cultures of umbilical cord mesenchymal stem cells in a hollow-fiber bioreactor show improved osteochondral regeneration activity. Cell Biol Toxicol 36:165-178 (2020). PubMed: 31820164
      • Zhu J  et al. Exosomes from nicotine-stimulated macrophages accelerate atherosclerosis through miR-21-3p/PTEN-mediated VSMC migration and proliferation. Theranostics 9:6901-6919 (2019). PubMed: 31660076
      • Anczurowski M  et al. Chaperones of the class I peptide-loading complex facilitate the constitutive presentation of endogenous antigens on HLA-DP84GGPM87. J Autoimmun 102:114-125 (2019). PubMed: 31078377
      • Moonschi FH  et al. Mammalian Cell-derived Vesicles for the Isolation of Organelle Specific Transmembrane Proteins to Conduct Single Molecule Studies. Bio Protoc 8:N/A (2018). PubMed: 30406159
      • Baba K  et al. Asiatic Acid, Corosolic Acid, and Maslinic Acid Interfere with Intracellular Trafficking and N-Linked Glycosylation of Intercellular Adhesion Molecule-1. Biol Pharm Bull 41:1757-1768 (2018). PubMed: 30504678
      • Fox-Loe AM  et al. Organelle-specific single-molecule imaging of a4ß2 nicotinic receptors reveals the effect of nicotine on receptor assembly and cell-surface trafficking. J Biol Chem 292:21159-21169 (2017). PubMed: 29074617
      • Shwetha S  et al. HuR Displaces Polypyrimidine Tract Binding Protein To Facilitate La Binding to the 3' Untranslated Region and Enhances Hepatitis C Virus Replication. J Virol 89:11356-71 (2015). WB ; Human . PubMed: 26339049
      • Mitsuda S  et al. Ursolic acid, a natural pentacyclic triterpenoid, inhibits intracellular trafficking of proteins and induces accumulation of intercellular adhesion molecule-1 linked to high-mannose-type glycans in the endoplasmic reticulum. FEBS Open Bio 4:229-39 (2014). ICC/IF ; Human . PubMed: 24649404

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      Western blot abreview for Anti-Calnexin antibody [EPR3632]

       Inconclusive
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (catecholaminergic neuronal tumor)
      Gel Running Conditions
      Reduced Denaturing (NuPAGE 4-12% Bis-Tris, 15 wells)
      Loading amount
      10 µg
      Treatment
      Chronically infected with RML prion strain
      Specification
      catecholaminergic neuronal tumor
      Blocking step
      Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 4°C
      Read More

      Dr. Manjeet Kumar

      Verified customer

      Submitted Mar 15 2019

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