Recombinant
RabMAb

Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633)

Rabbit recombinant monoclonal Carcino Embryonic Antigen CEA antibody [EPCEAR7]. Validated in WB, IHC, ICC/IF and tested in Human. Cited in 5 publication(s).

Overview

  • Product name
    Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7]
    See all Carcino Embryonic Antigen CEA primary antibodies
  • Description
    Rabbit monoclonal [EPCEAR7] to Carcino Embryonic Antigen CEA
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Human Carcino Embryonic Antigen CEA.

  • Positive control
    • Human colon cancer lysate, Human colon carcinoma tissue, MCF-7 and human gastric carcinoma.
  • 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.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133633 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 1/1000 - 1/10000. Detects a band of approximately 180-200 kDa (predicted molecular weight: 76 kDa).
IHC-P 1/3000.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

ICC/IF 1/50 - 1/500.

Target

  • Function
    Cell surface glycoprotein that plays a role in cell adhesion and in intracellular signaling. Receptor for E.coli Dr adhesins.
  • Tissue specificity
    Found in adenocarcinomas of endodermally derived digestive system epithelium and fetal colon.
  • Sequence similarities
    Belongs to the immunoglobulin superfamily. CEA family.
    Contains 7 Ig-like (immunoglobulin-like) domains.
  • Post-translational
    modifications
    Complex immunoreactive glycoprotein with a MW of 180 kDa comprising 60% carbohydrate.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Carcinoembryonic antigen antibody
    • Carcinoembryonic antigen-related cell adhesion molecule 5 antibody
    • CD66e antibody
    • CEA antibody
    • Ceacam5 antibody
    • CEAM5_HUMAN antibody
    • DKFZp781M2392 antibody
    • Meconium antigen 100 antibody
    • OTTHUMP00000199032 antibody
    • OTTHUMP00000199033 antibody
    • OTTHUMP00000199034 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1:3000 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1:500 dilution (3.6μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunocytochemistry/Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma cell line) labelling Carcino Embryonic Antigen CEA with purified ab133633 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human gastric carcinoma tissue sections labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1:3000 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • ab133633 showing positive staining in Normal colon tissue.

  • Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 1/1000 dilution + Human colon cancer lysate at 10 µg

    Secondary
    Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 76 kDa
    Observed band size: 180-200 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin embedded Human colon carcinoma tissue labelling Carcino Embryonic Antigen CEA with ab133633 at 1/250.
  • Anti-Carcino Embryonic Antigen CEA antibody [EPCEAR7] (ab133633) at 0.4 µg/ml (Purified) + Human colon cancer lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 76 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • ab133633 showing positive staining in Stomach adenocarcinoma tissue.

  • ab133633 showing negative staining in Skeletal muscle tissue.

  • ab133633 showing negative staining in Normal tonsil tissue.

  • ab133633 showing negative staining in Thyroid gland carcinoma tissue.

  • ab133633 showing negative staining in Hepatocellular carcinoma tissue.

  • ab133633 showing negative staining in Glioma tissue.

References

This product has been referenced in:
  • Ramos-Gomes F  et al. Single- and two-photon imaging of human micrometastases and disseminated tumour cells with conjugates of nanobodies and quantum dots. Sci Rep 8:4595 (2018). WB . Read more (PubMed: 29545609) »
  • Gao J  et al. Contrast-enhanced magnetic resonance imaging with a novel nano-size contrast agent for the clinical diagnosis of patients with lung cancer. Exp Ther Med 15:5415-5421 (2018). Read more (PubMed: 29904421) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for the details of your protocol and the image. I think the protocol is correct, and assuming your samples contain CEA, you should see a clear distinct band. The question is where you will see a band. The protein is heavily glycosylated in most tissues, but we have data from other CEA antibodies showing staining of a band closer to the MW of the 77 kDa (before post-translational modifications) precursor. For an example, see the blot on the datasheet of ab97480:

Click here (or use the following: https://www.abcam.com/index.html?datasheet=97480).

If you like, I will be happy to replace it with another vial or a vial of a different CEA antibody. Though we have not received other complaints for ab133633, I suggest the latter option, in particular the mouse monoclonal ab105364.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=105364).

The following paper has western blot data for human colorectal cancer cells using this clone:

Gastroenterology. 2005 Jun;128(7):1796-804.
PMID: 15940614

http://www.ncbi.nlm.nih.gov/pubmed?term=15940614

I look forward to your reply.

Read More

Answer

Thank you for bringing this to our attention and for sending the blot image. Can you please tell me if you are using protease inhibitors when you harvest the samples? How are you blocking the membrane, and what do you dilute the antibody in?

Read More

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