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

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

    This product is a recombinant rabbit monoclonal antibody.

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, 9.85% Tris glycine, 50% Tissue culture supernatant
  • 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

    • Lane 1: Hap1 wildtype cell lysate (20 µg)

      Lane 2: CANX Hap1 knockout cell lysate (20 µg)

      Lane 3: HEK-293T wildtype cell lysate (20 µg)

      Lane 4: CANX HEK-293T knockout cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 90 kDa. Red - loading control, ab8245 observed at 37 kDa.

      ab92573 was shown to react with Calnexin in HEK-293T wildtype. Loss of signal was observed when knockout sample ab263805 was used. Wild-type and Calnexin knockout samples were subjected to SDS-PAGE. ab92573 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 20000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Calnexin knockout HAP1 cell lysate (20 µg)
      Lane 3: THP-1 cell lysate (20 µg)
      Lane 4: RAW 264.7 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 1/10,000 dilution for 1 hour 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
      All lanes : 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.

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

    • 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.

    References

    This product has been referenced in:

    • 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). Read more (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). Read more (PubMed: 30504678) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    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

    Dr. Manjeet Kumar

    Verified customer

    Submitted Mar 15 2019

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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