Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)

Reviews (5)Q&A (9)References (57)

Overview

  • Product name

    Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker
    See all N Cadherin primary antibodies

  • Description

    Mouse monoclonal [5D5] to N Cadherin - Intercellular Junction Marker

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, IHC-Fr, WB, IHC-P, ICC/IF, ELISAmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment corresponding to Human N Cadherin.

  • Positive control

    • A431, NIH-3T3, Hela, C6 and LNCap cells. Human lung cancer, colon cancer, ovarian cancer and mammary cancer tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab98952 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
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr 1/200.
WB 1/500 - 1/2000. Predicted molecular weight: 100 kDa.
IHC-P 1/200 - 1/1000.
ICC/IF 1/200 - 1/1000.
ELISA 1/10000.

Target

  • Function

    Cadherins are calcium dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. CDH2 may be involved in neuronal recognition mechanism. In hippocampal neurons, may regulate dendritic spine density.

  • Sequence similarities

    Contains 5 cadherin domains.

  • Cellular localization

    Cell membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • CADH2_HUMAN antibody
    • Cadherin 2 antibody
    • Cadherin 2 N cadherin neuronal antibody
    • Cadherin 2 type 1 antibody
    • Cadherin 2 type 1 N cadherin neuronal antibody
    • Cadherin 2, type 1, N-cadherin (neuronal) antibody
    • Cadherin-2 antibody
    • Cadherin2 antibody
    • Calcium dependent adhesion protein neuronal antibody
    • CD325 antibody
    • CD325 antigen antibody
    • CDH2 antibody
    • CDHN antibody
    • CDw325 antibody
    • CDw325 antigen antibody
    • N cadherin 1 antibody
    • N-cadherin antibody
    • NCAD antibody
    • Neural cadherin antibody
    • OTTHUMP00000066304 antibody
    • OTTHUMP00000067378 antibody
    see all

Images

  • Western blot - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Western blot - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    All lanes : Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952) at 1/500 dilution

    Lane 1 : A431 cell lysate
    Lane 2 : NIH-3T3 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : C6 cell lysate
    Lane 5 : LNCap cell lysate

    Lysates/proteins at 15 µl per lane.

    Secondary
    All lanes : HRP-Goat Anti-Mouse Igg (Fc) at 1/10000 dilution

    Predicted band size: 100 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)This image is courtesy of an anonymous Abreview

    ab98952 staining N Cadherin in human NSCLC tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with using HOPE technique and blocked for 5 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 1 hour at 25°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)This image is courtesy of an anonymous Abreview

    ab98952 staining N Cadherin in paraffin-embedded human lung cancer tissue by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with HOPE and blocked. Samples were incubated with primary antibody (1/200) for 1 hour at 25°C. A TRITC-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Flow Cytometry - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Flow Cytometry - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Overlay histogram showing HeLa cells stained with ab98952 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab98952, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunohistochemical analysis of paraffin-embedded Human lung cancer (1), colon cancer (2), ovarian cancer (3) and mammary cancer (4), using ab98952 at 1/200 dilution with DAB staining.
  • Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunofluorescence analysis of A431 cells using ab98952 at 1/200 dilution (green). Blue: DRAQ5 fluorescent DNA dye
  • Immunohistochemistry (Frozen sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
    Immunohistochemistry (Frozen sections) - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)This image is courtesy of an Abreview submitted by Dr Nam-Ho Kim
    ab98952 staining N Cadherin (orange) in Mouse E15.5 Brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/200 in PBST) for 12 hours at 4°C. An Alexa Fluor® 568-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:

  • Liao Z  et al. High PLK4 expression promotes tumor progression and induces epithelial-mesenchymal transition by regulating the Wnt/ß-catenin signaling pathway in colorectal cancer. Int J Oncol 54:479-490 (2019). Read more (PubMed: 30570110) »
  • Tang H  et al. MARCH5 overexpression contributes to tumor growth and metastasis and associates with poor survival in breast cancer. Cancer Manag Res 11:201-215 (2019). Read more (PubMed: 30636894) »
See all 57 Publications for this product

Publishing research using ab98952? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 14 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (embryonic mouse brain coronal cortical section)
Permeabilization
Yes - Triton X
Specification
embryonic mouse brain coronal cortical section
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Dr. Bhavin Shah

Verified customer

Submitted Oct 20 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (NSCLC)
Antigen retrieval step
None
Permeabilization
No
Specification
NSCLC
Blocking step
ZytoChem-Plus HRP Polymer-Kit Blocking solution as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
HOPE
Read More

Abcam user community

Verified customer

Submitted Apr 13 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (paraffin-embedded lung cancer tissue)
Permeabilization
No
Specification
paraffin-embedded lung cancer tissue
Blocking step
Zytomed Systems HRP Polymer Blocking solution as blocking agent for 5 minute(s) · Concentration: 100%
Fixative
HOPE
Read More

Abcam user community

Verified customer

Submitted Mar 18 2014

Question

I would like to know the concentration of your product ab98952 which is the Anti-N Cadherin antibody when it is shipped. I see that the total volume of the antibody is 100 ul.

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Answer


ab98952 is an ascites product. As it is not a purified antibody, an exact concentration has not been determined. In general however, the specific antibody concentration in a product such as this is 0.9 - 9mg/ml.

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Question

Name: Irene Appolloni

Inquiry: Dear Sir or Madam, I am planning to carry out a double immunofluorescence staining for N-cadherin and beta-catenin antigens. I have the AbCam rabbit-anti N-cadherin antibody ab12221 that is a very good antibody but it requires a methanol/acetone fixation of the cells that is not compatible with my anti beta-catenin. Moreover, I’d rather use mouse anti N-cadheirn antibody to easily perform double immunostainings with rabbit antibodies. I noticed that you have the N-cadherin antibody [5D5] (ab98952), that I hope could solve all my problems. I would like to ask you whether it is possible to have a free sample aliquot to test it on my cells. Best regards,

Read More
Answer

Thank you for contacting us.

I am pleased to provide the following immunogen information for ab98952 N Cadherin antibody [5D5] :

Immunogen sequence: ##########

I am sorry to confirm that permeabilization was required when it was tested for Flow Cytometry and ICC.

In order to aid your research we are happy to provide this information to you. However, this immunogen sequence is proprietary (commercially sensitive and confidential). Therefore, please do not share or publish this information. It is strictly for your own use.

Regarding samples, because we carry over 90,000 products, I am sorry to confirm it is regrettably not feasible for us to stock small sample sizes of our products to send on request.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our AbTrial program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information. Or you can find more information about this program at the following link:

www.abcam.com/abtrial

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact me again with any other questions.

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Question

I have compared the 2 N-cadherin antibodies ab12221 and ab98952 on three samples and attach the results. The samples are identical and were run on the same gel and blotted onto a PVDF membrane using BSA in PBST to block. The membrane was then cut into 2 and the sections incubated overnight with the appropriate antibody in BSA/PBST.

As you can see, the results are very different. Of particular concern is the difference in relative amounts of the full-length N-cadherin. Indeed the a b98952 shows very little full-length at all.

Could you please advise on what might be the problem?

Many thanks,

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Answer

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.
I look forward to receiving your reply.

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Question

From your website, there is another N-Cad mouse antibody cat# ab124397 that could be tried, but it haveno reviews and no rating. Alternatively I could also try your cytokeratin 14 mouse antibody cat # ab7800 (I'm trying to do double staining with K14 and NCAD, I have the rabbit version of both already). Because the cytokeratin 14 is highly rated, I would like to try this one if possible. Thank you again for your help.

Read More
Answer

Thanks for the reply.

I am send you a vial of ab7800, mouse monoclonal to cytokeratin 14. Hopefully you will be able to get the desired staining pattern with this antibody. Your new order number is 1132112.

If there is anything else I can help you with, please let me know.

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Question

I have received the N-Cadherin antibody and tested it on my samples. But the problem persists that the antibody is picking up something in the mouse mammary epithelial cells. N-Cad antibodies from Novus or Santa Cruz do not have this and the side-by-side comparison shows no staining in the same cells.
Any suggestion what should I do now?

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Answer

Thank you for letting me know.

I am sorry about the continued problems that you have been having with ab98952. I will this information onto the lab.

Based on the information you have provided, I believe the best option is to go ahead and either replace the antibody with a different antibody, against the same target, or refund the cost of the antibody.

Please let me know how you would like to proceed.

I look forward to your reply.

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Question

Thank you for your help, I did notice that the label on the vial sent me was printed skewed (but still shows it to be N-Cad)and I was bit worried. I would be happy to have a another vial, preferably another lot,from Abcam.

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Answer

Thank you for your reply.

I am sending you a new vial of ab98952 to try out. Your new order number (for your records only) is *
.

If you have any issues with the new vial, or if there is anything else I can help you with, please let me know.

.

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Question


Thank you for your help. I have attached the picture file for
N-Cadherin comparisons (Novus vs Abcam). I strongly suspect that
Abcam ab is picking up E-Cadherin instead of N-Cadherin because
E-Cadherin is expressed by the luminal cells in the center.

Read More
Answer

Thank you for contacting Abcam.

I have been in contact with the lab regarding the staining pattern that you are observing whilst using ab98952. Our WB assay shows that the antibody is positive against recombinant protein. DNA sequencing results are consistent with the immungen sequence of this antibody, which are amino acids: **so we believe the antibody is against its intended target. Also we do have customer feedback that shows the anticipated staining pattern using this antibody. Therefore I believe that it may just be an issue with the vial that you received and so I would be very happy to send another vial of the antibody, free of charge, to see if that resolves the issue.
If, you would prefer to have a refund, then I would be happy to do that instead.
Please let me know how you would like to continue and I will be happy to help resolve this issue.
I look forward to your reply.

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1-10 of 14 Abreviews or Q&A

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