Overview

  • Product name
    Anti-Calnexin antibody [EPR3632]
    See all Calnexin primary antibodies
  • Description
    Rabbit monoclonal [EPR3632] to Calnexin
  • 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 (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Calnexin aa 1-100.

  • Positive control
    • WB: HeLa, A431, SH-SY5Y and HepG2 cell lysates. IHC-P: Human tonsil tissue. ICC/IF: Wild-type HAP1 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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

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

Properties

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.
  • 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

    Anti-Calnexin antibody [EPR3632] images



    • Predicted band size : 90 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Calnexin knockout HAP1 cell lysate (20 µg)
      Lane 3: THP1 cell lysate (20 µg)
      Lane 4: Raw264.7 cell lysate (20 µg)
      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 1/10000 dilution for 1 h at room temperature before imaging.

    • 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).

    • 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
      standard HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 90 kDa
    • Immunohistochemical analysis of paraffin embedded Human tonsil tissue using ab92573 at a 1/100 dilution.


    • Predicted band size : 90 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Calnexin knockout HAP1 cell lysate (20 µg)

      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.

    • Immunocytochemistry/Immunofluorescence analysis of human lung carcinoma A549 cells labelling Calnexin with ab92573. Cells were fixed with 4% PFA in PBS for 15 minutes washed three times with PBS, and then washed twice with 25 mM glycine–PBS. The cells were treated with cold methanol and washed three times with 0.3% Triton X-100 in PBS for permeabilization. The fixed cells were blocked with 5% BSA in PBS overnight. An Alexa Fluor® 594-conjugated anti-rabbit was used as the secondary antibody.

    References for Anti-Calnexin antibody [EPR3632] (ab92573)

    This product has been referenced in:
    • 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 . Read more (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 . Read more (PubMed: 24649404) »

    See all 2 Publications for this product

    Product Wall

    There are currently no Abreviews or Questions for ab92573.
    Please use the links above to contact us or submit feedback about this product.

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

    Sign up