Overview

  • Product name
    Anti-pan Cadherin antibody
    See all pan Cadherin primary antibodies
  • Description
    Rabbit polyclonal to pan Cadherin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IF, Electron Microscopy, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, Xenopus laevis, Zebrafish
    Predicted to work with: Rabbit, Bird, Fish, Amphibian
  • Immunogen

    Synthetic peptide:

    DYDYLNDWGPRFKKLADMYGGGDD

    conjugated to KLH by a Glutaraldehyde linker, corresponding to C terminal amino acids 889-912 of Chicken pan Cadherin.

  • General notes
    Immunohistochemistry This antibody has not been tested on frozen sections but has been tested on both cultured cells and formalin-fixed, paraffin-embedded, protease digested sections so chances are it will work on frozen sections. It has also been tested on heart sections and MDBK cells. This antibody should work on bovine samples although we have not tested it specifically.


    This product is useful for the detection of members of the cadherin family or genetically engineered proteins containing the C-terminal cadherin tail, and for demonstration of adherens type cell-cell junctions regardless of their cadherin type.

Properties

Applications

Our Abpromise guarantee covers the use of ab6529 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 1/1000. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.
WB 1/200. On chicken and rabbit heart extract this antibody detects a single band at 135 kD.
ICC/IF 1/100. Fix cells with 2-4% paraformaldehyde for 10 minutes at room temperature, followed by 10-minute incubation with 0.2% Triton X-100 (see Gellersen et al 2007 and Kaur et al 2006).
Electron Microscopy Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. Fix with ice cold ethanol.

Target

Images

  • Immunofluorescent imaging of human cells (U2OS) with ab6529 confirms the specificity of this antibody.  Pan cadherin antibody recognises predominantly membranous signal with some fainter cytoplasmic staining corresponding to soluble cadherin. No nuclear staining is seen. 

    IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes.  All blocking and incubation steps carried out at 37 degrees C. Nuclei were stained with Hoechst stain.

  • ab6529 staining pan Cadherin in human bone marrow cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed and blocked in 2% BSA for 30 minutes at 20°C. The primary antibody was diluted, 1/200 and incubated with sample for 9 hour at 4°C. An Alexa Fluor® 488 conjugated goat polyclonal to rabbit IgG, diluted 1/200 was used as secondary.

  • All Lanes: pan Cadherin antibody (ab6529) at 1/500 dilution + Mouse brain whole tissue lysate (100 µg)

    Secondary Antibody: An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal developed using the ECL technique

    Blocking Step: 5% Milk for 1 hour at 25 °C.

    Gel Running Conditions: Reduced, denaturing.

    See Abreview

  • ab6529 staining pan Cadherin in rat kidney tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with acetone and then blocked with 2% BSA for 2 hours at 25°C followed by incubation with the primary antibody, at a 1/200 dilution, for 9 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 (green) used at a 1/500 dilution.

    See Abreview

  • ab6529 staining pan Cadherin in murine liver cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in formaldehyde, permeabilised in 0.4% Triton X-100 and then blocked using 10% serum for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/200 for 17 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 594 (red) used at a 1/1000 dilution. The other secondary antibody used was an Alexa Fluor® 488 (green) goat anti-mouse IgG for detection of a specific cytosolic protein.

    See Abreview

References

This product has been referenced in:
  • Choi EK  et al. Functional analysis of SLC39A8 mutations and their implications for manganese deficiency and mitochondrial disorders. Sci Rep 8:3163 (2018). Read more (PubMed: 29453449) »
  • Higgins SJ  et al. Tie2 protects the vasculature against thrombus formation in systemic inflammation. J Clin Invest 128:1471-1484 (2018). Read more (PubMed: 29360642) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 31 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Roti­-ImmunoBlock as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 26°C
Sample
Cat Cell (CrFK)
Specification
CrFK
Permeabilization
Yes - Triton X-100 0.2 %
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 27 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Liver (20 um section))
Specification
Liver (20 um section)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 18 2013

Answer

Merci de nous avoir contactés.

Comme convenu, j'ai demandé à notre service comptabilité de mettre en place un avoir référence CN.

Selon nos archives, vous avez désormais deux avoirs à utiliser :

Référence CN xxxxx d’une valeur de €xxxxx (contre la commande 971341)

Référence CN xxxxxxx d’une valeur de €xxxxx (contre la commande 1164385)

Voici les manières dont vous pouvez utiliser ces références CN:
(1) les faire valoir contre la facture originale si celle-ci n'as pas encore été payée
(2) les faire valoir contre une prochaine facture
(3) obtenir un remboursement de la somme entière si vous n'avez pas de factures actuellement en cours avec Abcam

Si vous souhaitez recevoir un remboursement au lieu d'un avoir, veuillez demander à votre service de gestion de prendre contact avec notre département comptabilité afin que nous puissions recueillir les informations nécessaires pour cette restitution. Notre service comptabilité peut être contacté par courriel à l'adresse creditcontrol@abcam.com ou par téléphone en utilisant le lien « Contactez-nous » dans le coin en haut à droite de notre site internet. Veuillez communiquer votre numéro d'avoir lorsque vous contactez notre service comptabilité.

Vous recevrez également la référence complète de la note de crédit par courriel ou par voie postale; cette référence commençant par les lettres CGB.

Bonne continuation dans votre recherche.

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Answer

Merci de nous avoir contactés.

Je suis désolée de n'avoir pu améliorer vos résultats.

Je vous propose un remboursement ou un avoir. Sinon je peux également vous proposer un remplacement avec l’anticorps ab7671, Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] - Plasma Membrane Marker (https://www.abcam.com/ab7671) qui pourrait marcher.

Si vous pouvez me faire savoir quelle résolution vous préférez.

J'espère que cette expérience ne vous empêchera pas de commander à nouveau l'un de nos produits.

L'équipe technique reste à votre disposition pour plus de conseils.

Read More

Answer

Merci d'avoir pris le temps de nous envoyer votre protocole et images. Je suis désolée d'apprendre que vous avez eu des difficultés à obtenir de bons résultats avec ab6529

Notre politique est d'offrir tout d'abord la meilleure assistance technique. Si nous concluons que le produit que vous avez reçu ne fonctionne pas comme mentionné sur sa fiche technique et que ce produit a été commandé sous 120 jours, nous nous ferons un plaisir de vous offrir un remplacement gratuit ou un avoir en compensation.

J'aimerais donc vous offrir quelques suggestions afin d'optimiser vos résultats:

Premièrement, il me semble que il y’a peut-être trop de perméabilisations avec le triton, car il est utiliser dans toutes les étapes de blocages et d’incubations. Puisque triton est assez fort comme agent perméabilisant, cela pourrait peut-être dégrader les membranes et donc affecter le marquage. Je vous suggère donc deux choses :

Vous pouvez avoir une étape de perméabilisation après l'étape de fixation en utilisant 0,1-0,4% triton pendant 5-10 minutes puis incuber avec les anticorps (sans ajouter de triton)
Vous pouvez également changer l’agent perméabilisant à un qui soit plus doux, par exemple 0,2% tween et l’ajouter a l’étape du blocage



Sinon il y’a d’autre suggestions que je peux vous proposer:

Essayer un fixateur différent par exemple éthanol glacé pendant 5 minutes
Augmenter la concentration du sérum en tant qu’agent bloquant à 10% pour une incubation d’une heure pour réduire les marquages non-spécifiques
Diluer l’anticorps primaire à 1:200 1:400




J'espère que ces informations vous aideront. Si nos suggestions ne permettent pas d'améliorer vos résultats, n'hésitez pas à me recontacter pour plus d'assistance.

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Question
Answer

Je suis désolée d'apprendre que ab6529 ne vous donne pas de bons résultats en IF.

Veuillez trouver ci-joint un questionnaire qui nous permet de regrouper le plus d'informations possible sur le protocole que vous avez utilisé avec cet anticorps.
Celui-ci nous permettra de vous fournir la meilleure assistance technique.

Si nous concluons que notre produit est fautif et ne fonctionne pas comme explicité par notre fiche technique, nous serons ravis de mettre en place l'envoi d'un tube de remplacement gratuit ou d'un remboursement (sous réserve d'une commande ultérieure de 120 jours).

Je vous remercie par avance de nous envoyer votre protocole.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepG2)
Specification
HepG2
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 29 2012

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.

Read More

Answer

Thank you for the images of the staining with ab22744.

I agree the staining is not what is usually observed for cadherin. I think you are expecting something like what one of our reviewers saw. The data is at this link:

https://www.abcam.com/index.html?datasheet=22744&tab=abreviews&intabreviewid=22768

Are you still fixing your cells with PFA? It may be that correct staining is dependent on the fixation protocol. The data we have for ab22744 is all from methanol-fixed cells. I suggest trying this if you are able to, fixation in cold methanol for five minutes, followed by rinsing with PBS.

Please let me know if you are willing to try this. If not, we can discuss replacement with a Na ATPase antibody.

Read More

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab22744.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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1-10 of 31 Abreviews or Q&A

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