Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (HRP) (ab195198)

Overview

  • Product name

    Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (HRP)
    See all Calnexin primary antibodies
  • Description

    Rabbit monoclonal [EPR3633(2)] to Calnexin - ER Membrane Marker (HRP)
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

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

    Reacts with: Human
    Does not react with: Mouse
  • Immunogen

    Synthetic peptide within Human Calnexin aa 550 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, A431 and HeLa whole cell lysates. IHC-P: normal human colon tissue.
  • General notes

    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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% Proclin
    Constituents: 1% BSA, 30% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3633(2)
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab195198 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 1/5000. Detects a band of approximately 82 kDa (predicted molecular weight: 68 kDa).
IHC-P Use at an assay dependent concentration.

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

  • All lanes : Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (HRP) (ab195198) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : CANX (Calnexin) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 68 kDa



    ab195198 was shown to recognize Calnexin in wild-type HAP1 cells as signal was lost at the expected MW in CANX (Calnexin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CANX (Calnexin) knockout samples were subjected to SDS-PAGE. Ab195198 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • IHC image of Calnexin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab195198 at 1/500 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-Calnexin antibody [EPR3633(2)] - ER Membrane Marker (HRP) (ab195198) at 1/5000 dilution

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HeLa whole cell lysate (ab150035)

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 82 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195198 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

This product has been referenced in:

  • Tang C  et al. Aspartate ß-hydroxylase disrupts mitochondrial DNA stability and function in hepatocellular carcinoma. Oncogenesis 6:e362 (2017). Read more (PubMed: 28714949) »
See 1 Publication for this product

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