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    n-cadherin-antibody-5d5-intercellular-junction-marker-ab98952.pdf

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Signal Transduction Cytoskeleton / ECM Cell Adhesion Cadherins
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Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)

  • Datasheet
  • SDS
Reviews (5)Q&A (9)References (78)

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Western blot - 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)
  • Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)
  • Flow Cytometry - 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)
  • Immunocytochemistry/ Immunofluorescence - Anti-N Cadherin antibody [5D5] - Intercellular Junction Marker (ab98952)

Key features and details

  • Mouse monoclonal [5D5] to N Cadherin - Intercellular Junction Marker
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

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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 & Species

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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    Purified from tissue culture supernatant.
  • Clonality

    Monoclonal
  • Clone number

    5D5
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Cadherins
    • Stem Cells
    • Hematopoietic Progenitors
    • Surface Molecules
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Cadherins

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Positive Controls

    • HeLa membrane extract lysate (ab29547)
    • A-431 membrane extract lysate (ab30134)
  • Recombinant Protein

    • Recombinant Human N Cadherin protein (His tag) (ab223009)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
Flow Cyt
1/100.

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

WB
1/500 - 1/2000. Predicted molecular weight: 100 kDa.
IHC-P (1)
1/200 - 1/1000.
ICC/IF (2)
1/200 - 1/1000.
Notes
Flow Cyt
1/100.

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

WB
1/500 - 1/2000. Predicted molecular weight: 100 kDa.
IHC-P
1/200 - 1/1000.
ICC/IF
1/200 - 1/1000.

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.
  • Target information above from: UniProt accession P19022 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1000 Human
    • Entrez Gene: 12558 Mouse
    • Entrez Gene: 83501 Rat
    • Omim: 114020 Human
    • SwissProt: P19022 Human
    • SwissProt: P15116 Mouse
    • SwissProt: Q9Z1Y3 Rat
    • Unigene: 464829 Human
    • Unigene: 606106 Human
    • Unigene: 257437 Mouse
    • Unigene: 23200 Rat
    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

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (78)

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

    ab98952 has been referenced in 78 publications.

    • Chow T  et al. Recapitulating kidney development in vitro by priming and differentiating mouse embryonic stem cells in monolayers. NPJ Regen Med 5:7 (2020). PubMed: 32351711
    • Zheng Y  et al. Altered expression of glycoprotein non-metastatic melanoma protein B in the distal sciatic nerve following injury. Int J Mol Med 45:1909-1917 (2020). PubMed: 32236569
    • Tan S  et al. RAC1 Involves in the Radioresistance by Mediating Epithelial-Mesenchymal Transition in Lung Cancer. Front Oncol 10:649 (2020). PubMed: 32411607
    • Tiwari R  et al. Androgen deprivation upregulates SPINK1 expression and potentiates cellular plasticity in prostate cancer. Nat Commun 11:384 (2020). PubMed: 31959826
    • Yang L  et al. SP1 induced long non-coding RNA LINC00958 overexpression facilitate cell proliferation, migration and invasion in lung adenocarcinoma via mediating miR-625-5p/CPSF7 axis. Cancer Cell Int 20:24 (2020). PubMed: 31997940
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 14 Abreviews or Q&A

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

    Excellent
    Abreviews
    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
    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
    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.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 20 2013

    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.

    Read More

    Abcam Scientific Support

    Answered on Feb 20 2013

    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

    Abcam community

    Verified customer

    Asked on Dec 05 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Dec 05 2012

    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,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 25 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 25 2012

    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

    Abcam community

    Verified customer

    Asked on Jul 23 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 23 2012

    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?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 20 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 20 2012

    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.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 16 2012

    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.

    .

    Read More

    Abcam Scientific Support

    Answered on Jul 16 2012

    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

    Abcam community

    Verified customer

    Asked on Jul 16 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 16 2012

    1-10 of 14 Abreviews or Q&A

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