Astana Biomed Group, an authorized Abcam distributor in Central Asia

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: Mouse, 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 & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa, A431, SH-SY5Y, HEK-293T, MCF7, U-2 OS 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.

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

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 HEK-293T cell lysate
    Lane 2 : CANX knockout HEK-293T cell lysate
    Lane 3 : U-2 OS cell lysate
    Lane 4 : MCF7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 90 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab92573 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab92573 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 20000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h 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 : 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.

  • 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 rabbit polyclonal antibody.

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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