Anti-N Cadherin antibody (ab18203)

Rabbit polyclonal N Cadherin antibody. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human, Pig, Xenopus laevis. Cited in 153 publication(s).

Overview

  • Product name
  • Description
    Rabbit polyclonal to N Cadherin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, WB, IHC-P, IP, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig, Xenopus laevis
    Predicted to work with: Chicken, Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human N Cadherin.

    Read Abcam's proprietary immunogen policy (Peptide available as ab18620.)

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Rat Brain Normal, Mouse Brain Normal, Human Brain Normal. This antibody gave a positive signal in the following tissues: Formalin Fixed Paraffin Embedded Human Liver Carcinoma. ICC/IF: HaCaT cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab18203 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-Fr 1/300.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 125-135 kDa (predicted molecular weight: 100 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use a concentration of 10 µg/ml.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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
  • 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

  • All lanes : Anti-N Cadherin antibody (ab18203) at 1 µg/ml

    Lane 1 : Brain (Rat) Tissue Lysate at 10 µg
    Lane 2 : Brain (Mouse) Tissue Lysate at 10 µg
    Lane 3 : Brain (Human) Tissue Lysate at 20 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 100 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    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 2% Bovine Serum Albumin before being incubated with ab18203 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    The N Cadherin protein has a predicted molecular weight of 100 kDa, however it is extensively glycosylated and has been shown to run in the 125-135 kDa region (SwissProt data).

  • Anti N-cadherin (ab18203) staining of mouse brain using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London.

  • Anti N-cadherin (ab18203) staining of human ovarian cancer tissue using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London

  • Anti N-cadherin (ab18203) staining in a human melanoma xenograft mouse model using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London.

  • Anti N-cadherin (ab18203) staining of E17 developing rat retina using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/1000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London.

  • IHC image of N Cadherin staining in Human liver cancer 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 ab18203, 1µg/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.

  • anti-N cadherin antibody ab18203 stained human embryonic stem cells differentiated into mesoderm.
  • N Cadherin was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to N Cadherin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18203.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 135kDa: N Cadherin
  • ab18203 staining CD63 in human Fibrosarcoma HT1080 cell line by flow cytometry. Cells were incubated with primary antibody for 1 hour. ab7007, a donkey polyclonal to rabbit Ig, was used as the secondary antibody.

    See Abreview

  • ab18203 staining N Cadherin in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a 10 mM citrate buffer pH6.0. Samples were incubated with primary antibody (1/100 in PBS plus casein) for 90 minutes at 37°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Paraformaldehyde-fixed, 0.2% Triton X100 permeabilized HaCaT (human keratinocyte cell line) cells stained for N Cadherin (green) using ab18203 at 1/200 dilution in ICC/IF, followed by Donkey anti Rabbit Alexa Fluor 568 at 1/500 dilution.

    See Abreview

  • Immunohistochemistry of kidney carcinoma staining N Cadherin with ab18203 at 1μg/ml

References

This product has been referenced in:
  • Xu XF  et al. Suppression of BMX-ARHGAP fusion gene inhibits epithelial-mesenchymal transition in gastric cancer cells via RhoA-mediated blockade of JAK/STAT axis. J Cell Biochem 120:439-451 (2019). Read more (PubMed: 30216523) »
  • Gao J  et al. MicroRNA-590-5p functions as a tumor suppressor in breast cancer conferring inhibitory effects on cell migration, invasion, and epithelial-mesenchymal transition by downregulating the Wnt-ß-catenin signaling pathway. J Cell Physiol 234:1827-1841 (2019). Read more (PubMed: 30191949) »
See all 160 Publications for this product

Customer reviews and Q&As

1-6 of 6 Q&A

Question
Answer



ab18203 should be stored at -20 or -80 degrees C for long term storage. If stored in the freezer in single use aliquots, its expected this antibody will be stable for 3-5 years.

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Answer

Thank you for contacting us.

I've had a look through what information we have and unfortunately it doesn't look like we have done any additional testing to confirm whether the bands at 50kDa and 25kDa on the WB for ab18203 are non-specific or a truncated form of the protein.

I am sorry that on this occasion I couldn't be more helpful. 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

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

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 testing discount 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.

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 us again with any other questions.

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Answer

Thank you for your inquiry.

I can confirm that the immunogen is a synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human N Cadherin.

According to the UniProt homepage (http://www.uniprot.org/uniprot/P19022) amino acids 746-906 are cytoplasmic.

I can therefore confirm that a permeabilisation will be necessary for your experiments.

I wish you good luck and hope this information was helpful.

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Answer

Thank you for contacting Abcam.

I am glad to hear that the antibody is working well in bovine tissues. I was wondering if you would be able to provide me with some additional information about your negative controls, is control IgG an Abcam product? Does it match the species, isotype, clonality and conjugation of ab18203. Have you considered using the blocking peptide that is available as a negative control (ab18620; https://www.abcam.com/n-cadherin-peptide-ab18620.html)?
Ab18203 is not tested by us for specific reaction with bovine tissue. The "predicted to work" classification is based on homology between the immunogen and the bovine protein.

Please let me know if you have any questions.

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Answer

Thank you and your customer for your inquiry. According to the SwissProt homepage the cytoplasmic domain is only predicted to be from 746 to 900 amino acids. To my knowledge there is no experimental data to confirm this. If your customer would like test if the antibody sees the cytoplasmic domain even in un-fixed and un-permeabilised cells I suggest to run a control with fixed and permeabilised cells: 1. control: fix and perm and then stain 2. stain and then fix and perm. Both samples have to be fixed and permeabilised since fixed cell look different in the forward/sideward scatter. I hope this information is helpful for your customer. Please do not hesitate to contact me again with any further questions.

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