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Signal Transduction Cytoskeleton / ECM Cell Adhesion Cadherins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
  • Flow Cytometry - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
  • Immunoprecipitation - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
  • Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22397-264] to N Cadherin - BSA and Azide free
  • Suitable for: WB, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Conjugation
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PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
Protein
Product image
Recombinant Human N Cadherin protein (His tag) (ab223009)

View more associated products

Overview

  • Product name

    Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free
    See all N Cadherin primary antibodies
  • Description

    Rabbit monoclonal [EPR22397-264] to N Cadherin - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, IPmore details
    Unsuitable for: ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human N Cadherin aa 150-750. The exact sequence is proprietary.
    Database link: P19022

  • Positive control

    • WB: HeLa, PC-3, C6, A549, HEK-293T and HepG2 whole cell lysate. Human brain lysate. Mouse brain and heart lysate. Rat brain, heart and liver lysate. Flow Cyt: MCF7 cells. IP: N Cadherin IP in HeLa whole cell lysate.
  • General notes

    Ab245827 is the carrier-free version of ab245117. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab245827 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22397-264
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-N Cadherin antibody [EPR22397-264] (ab245117)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

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

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CDH2 (N Cadherin) knockout HEK293T cell line (ab255377)
  • KO cell lysates

    • Human CDH2 (N Cadherin) knockout HEK293T cell lysate (ab263843)
  • Recombinant Protein

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

Applications

Our Abpromise guarantee covers the use of ab245827 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 Use at an assay dependent concentration. Detects a band of approximately 130, 110 kDa (predicted molecular weight: 100 kDa).
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.
  • 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 [EPR22397-264] - BSA and Azide free (ab245827)
      Western blot - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
      All lanes : Anti-N Cadherin antibody [EPR22397-264] (ab245117) at 1/100000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : CDH2 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 100 kDa
      Observed band size: 130 kDa
      why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab245117).

      Lanes 1 - 2: Merged signal (red and green). Green - ab245117 observed at 125 kDa. Red - loading control, ab8245 observed at 37 kDa.  

       ab245117 was shown to react with N Cadherin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255377 (knockout cell lysate ab263843) was used. Wild-type and N Cadherin knockout samples were subjected to SDS-PAGE. ab245117 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Flow Cytometry - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
      Flow Cytometry - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)

      Flow cytometric analysis of MCF7 (Human breast adenocarcinoma epithelial cell, Left) / HeLa (Human cervix adenocarcinoma epithelial cell, Right) cell lines labeling N Cadherin with ab245117 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.


      Negative control: MCF7 (PMID: 9177902).


      Gated on viable cells.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245117).
    • Immunoprecipitation - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
      Immunoprecipitation - Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)

      N Cadherin was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell line) whole cell lysate with ab245117 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245117 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: HeLa whole cell lysate 10 µg (Input).
      Lane 2: ab245117 IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245117 in HeLa whole cell lysate.
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 seconds.

      The molecular weight is consistent with literature (PMID: 8230319).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245117).

    • Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)
      Anti-N Cadherin antibody [EPR22397-264] - BSA and Azide free (ab245827)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab245827? Please let us know so that we can cite the reference in this datasheet.

    ab245827 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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