Overview

  • Product name
    Anti-Carcino Embryonic Antigen CEA antibody [26/3/13]
    See all Carcino Embryonic Antigen CEA primary antibodies
  • Description
    Mouse monoclonal [26/3/13] to Carcino Embryonic Antigen CEA
  • Host species
    Mouse
  • Specificity
    This antibody recognises exclusively CEACAM 5 transiently expressed on the cell surface of transfected BOSC cells. It can be used to distinguish CEACAM 5 from all other CEACAM and probably all pregnancy-specific glycoproteins molecules, namely CEACAM 1 (BGP/CD66a), CEACAM 3 (CGM1/CD66d), CEACAM 4 (CGM7), CEACAM 6 (NCA/CD66c), CEACAM 7 (CGM2), CEACAM 8 (CGM6/CD66b)and PSG1 (CD66f) based on its reactivity pattern with stable HeLa transfectants expressing individual CEA family members. This antibody was included and characterized in the studies from the VIth Leucocyte Typing Workshop (Grunert F et al. 1994)
  • Tested applications
    Suitable for: IHC-P, Sandwich ELISA, ELISA, Flow Cyt, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length native protein (partially purified) (Human) from a perchloric acid extract from liver metastases of colonic tumors (Grunert F, et al. 1985)

  • Positive control
    • IHC-P: human normal colon tissue sections
  • General notes
    Antibodies produced from cDNA: Conventional technologies usually either generate antibodies against purified proteins, or against synthetic peptides based on amino acid sequences derived from DNA sequence data. Genetic immunization involves introducing the gene in the form of a cDNA directly into an animal which translates this cDNA into protein thus stimulating an immune response against the foreign protein. Although the synthetic peptide approach is comparable in speed, the quality of antibodies generated by genetic immunization is far superior. This is because the protein is made by the immunized animal, utilzing complex cellular mechanisms that allow it to gain a native conformation. Antibodies are then generated against a native protein, such as is found in the blood or tissues of its host species. Membrane-bound or secreted proteins often create problems for conventional antibody technology because in their native form, they are often modified by glycosylation, or in some cases exist as multiple membrane-spanning proteins that are not soluble following isolation or synthesis in recombinant systems. All of these problems are avoided if the immunized animal makes the protein itself. Antibodies generated by genetic immunization have been shown to have binding affinities to the protein in the sub-nanomolar range, which are approximately 100x higher than conventionally developed antibodies and much higher than single chain antibodies. Results confirm published data for much higher avidity of sera generated by genetic immunization as compared with that gained by immunization with a corresponding recombinant protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab4451 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Sandwich ELISA Use a concentration of 0.2 µg/ml.

For sandwich ELISA, use this antibody as Capture at 0.2µg/ml

ELISA Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

WB Use at an assay dependent concentration. Predicted molecular weight: 84 kDa.
IHC-Fr Use at an assay dependent concentration.

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

  • IHC image of Carcino Embryonic Antigen CEA staining in human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab4451, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    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.
  • Standard Curve for Carcino Embryonic Antigen CEA dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [26/3/13] to Carcino Embryonic Antigen CEA (ab4451) at 0.2ug/ml and Detector Antibody Rabbit polyclonal to Carcino Embryonic Antigen CEA (ab15987) at 0.5ug/ml

References

This product has been referenced in:
  • Li N  et al. Contrast-enhanced ultrasound with a novel nanoparticle contrast agent for clinical diagnosis in patients with non-small cell lung cancer. Exp Ther Med 14:3768-3773 (2017). Read more (PubMed: 29042977) »
  • Stern LA  et al. Geometry and expression enhance enrichment of functional yeast-displayed ligands via cell panning. Biotechnol Bioeng 113:2328-41 (2016). Read more (PubMed: 27144954) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

Thank you for your email.
Yes, that is correct. Alternatively, you can just use a secondary antibody that has already been conjugated with HRP. We carry many in our catalog. Please let me know if you need help selecting one.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

The conjugation kit ab102891 is compatible with antibody formulations that contain up to 1% BSA, even though the BSA is in 10-fold excess to the antibody (10mg/ml BSA compared to 1 mg/ml IgG for ab71720). Some of the BSA will itself will be conjugated, affecting efficient conjugation of the IgG, so an antibody without BSA would be better, such as anti-PSMA ab19071.

However, even with 1% BSA, much of the IgG in ab71720 can be conjugated to HRP using the HRP kit ab102891, and will be useful for subsequent assays. I do not know if that is the case for the other company's conjugation kit.

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Answer

I think the CEA antibody formulation will be compatible with the other company's labeling kit but you should contact them and send them the buffer components. Here are the components of the ab15987 buffer:

Preservative: 15mM Sodium Azide Constituents: 0.1M Sodium chloride, pH 7.2

The concentration of IgG for our current lot of ab15987 is 2.0 mg/ml. There is no protein carrier such as BSA.

The PSMA antibody formulation (ab97080) may be more problematic, as it contains BSA, which may interfere with conjugation of the HRP to the immunoglobulin. The buffer components are:

Preservative: 0.02% Sodium Azide Constituents: 1% BSA, PBS, pH 7.4

The IgG concentration of ab97080 is 1.0 mg/ml.

Again, the other company will be able to tell you if these components are compatible with their conjugation kit. You may need to remove the BSA, in which case I recommend our purification product, ab102784.

https://www.abcam.com/index.html?datasheet=102784

We also have an HRP conjugation kit, if you are interested. The catalogue number is ab102891.

https://www.abcam.com/index.html?datasheet=102891

I hope this is helpful. If you have any questions, please contact us.

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Answer

Thank you for your resposne.

As my colleauge has explained in the previous e-mail, our antibodies and proteins will regrettably not have been tested and guaranteed in the application you are using (i.e. electrical biosensors).

As ELISA is the closest application, I am pleased to recommend some antibodies suitable for this applications, however these will not be covered by our guarantee for electrical biosensors.

CA 15.3 - none

TRU-QUANT - none

CA 27.29 - none

CA125 - ab10033, ab10029, ab19551

CEA (carcinoembryonic antigen) - ab15987, ab4451,



If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Thank you for your enquiry.

I can confirm that ab4451 and ab15987 have been tested and guarantee for use in sELISA together according to the datasheets. Theab742 Carcino Embryonic Antigen CEA protein is full length native protein so should be suitable as a standard for use with this antibodies.

I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Question

Dear Sir:
Thank you for your kind reply. My samples are human derived (human blood and serum or urine). I am interested in Sandwich ELISA for Carcinoembryonic Antigen and Prostate-specific membrane antigen.
I am waiting for your suggestion on primary, secondary antibodies and antigenfor the samewhich are available with you.
Regards
Sunil Bhand

On Thu, Mar 1, 2012 at 5:52 AM, <mailto:technical@abcam.com> wrote:



Dear Dr Bhand,

We have an answer to your inquiry:
Thank you for contacting us. I will be happy to make some suggestions. Can you please confirm that your samples are human-derived, and that you are interested in a sandwich ELISA, rather than direct or indirect ELISA?

For an example of a sandwich ELISA antibody pair that we recommend, two antibodies that have been tested together for detection of human CEA by sandwich ELISA are ab4451 (for capture onto the plate) and ab15987 (for detection). The rabbit polyclonal ab15987 is unconjugated, so you will need to detect it in turn with an anti-rabbit IgG antibody, such as the HRP conjugate ab97080.

For the CEA antigen, and for the PSA assay, we have several choices but I will wait for your information regarding your samples and assay requirements before making suggestions.
Help us improve our service. Rate your experience with us today. https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=3557625

Your original inquiry to Abcam:
-------------------------------------------

I want to perform ELISA for Carcinoembryonic Antigen and Prostate-specific membrane antigen. Please suggest me the suitable primary as well as secondary conjugated antibodies for both. Also I want to procure the said antigens and antibodies.
Kindly send me the quotations for the above mentioned products.

-------------------------------------------


Best regards,
Thomas

Thomas Ruyle
Scientific Support Specialist
Abcam Inc.
https://www.abcam.com


Abcam Customer Services and Technical Support Team

https://www.abcam.com/technical
[CCE3557625]

--
Thanks & Regards
Deepti Parab
Office Assistant to
Dr. Sunil Bhand
Department of Chemistry
NAIP Project, BITS Pilani-K.K.Birla Goa Campus
Contact : 0832 2580340

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Answer

For the CEA assay, I recommend the antibodies I mentioned in my previous e-mail, ab4451 for capture and ab15987 for detection, followed by the secondary, anti-rabbit IgG conjugated to HRP, ab97080, and appropriate substrate, for instance, TMB.
For CEA antigen, I recommend ab724:

https://www.abcam.com/index.html?datasheet=724

For the PSMA assay, we do not have a pair of antibodies that has been tested together in sandwich ELISA. However, the following two antibodies are likely to be an appropriate pair, as they are raised against different portions of the PSMA protein: ab19071 for capture, and ab71720 for detection, followed by ab97080, the rabbit IgG-specific HRP conjugate, and substrate.

However, antibody ab71720 has not been tested in ELISA, and I cannot guarantee it will work well in this application, though the western blot looks very good.

We do not have an appropriate PSMA protein in our catalogue, so I suggest searching the website www.biocompare.com.

I hope this helps. If you have any questions, please contact me. For prices, please consult the online datasheets.

Read More

Answer

Thank you for contacting us. I will be happy to make some suggestions. Can you please confirm that your samples are human-derived, and that you are interested in a sandwich ELISA, rather than direct or indirect ELISA?

For an example of a sandwich ELISA antibody pair that we recommend, two antibodies that have been tested together for detection of human CEA by sandwich ELISA are ab4451 (for capture onto the plate) and ab15987 (for detection). The rabbit polyclonal ab15987 is unconjugated, so you will need to detect it in turn with an anti-rabbit IgG antibody, such as the HRP conjugate ab97080.

For the CEA antigen, and for the PSA assay, we have several choices but I will wait for your information regarding your samples and assay requirements before making suggestions.

Read More

Question
Answer

Thank you for your inquiry.


You are inquiring about the sELISA pair, ab4451 for capture and ab15987 for detection, correct?


Currently we do not offer a biotinylated form of ab15987. If you are looking to order this antibody in bulk (> 10 vials) we may be able to see if the lab can make a custom preparation. Alternatively, we do sell conjugation kits, which when used with our concentration kit to remove the azide from ab15987, would allow you to directly biotinylate ab15987 in 3 steps.


We sell the conentration kits as 1 column (ab102776) or 3 column (ab102778).


https://www.abcam.com/Antibody-Concentration-Kit-1-column-ab102776.html

https://www.abcam.com/Antibody-Concentration-Kit-3-columns-ab102778.html


We sell the biotinylation kit in different sizes, but I've included the 3 x10ug size below:


https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-3-x-10µg-Biotin-ab102865.html


I hope this information helps. Please contact us with any other questions.

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Application
ELISA
Sample
Human Purified protein (not sure)
Specification
not sure
Type
Sandwich (Capture)

Abcam user community

Verified customer

Submitted Jan 11 2011

Answer

Thank you for your enquiry. CEACAM5 [26/3/13]ab4451 belongs to the GOLD1 inhibition group. However, it does not inhibit all other GOLD1 members, and it does not bind to the isolated N-terminal domain of CEACAM5 if expressed alone. Unfortunately, this antibody has not been epitope mapped. Therefore we do not know the exact peptide sequence that the antibody reacts to.

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