Anti-Claudin 1 抗体 (ab15098)

According to our records, ab15098 and ab53765 were proving difficult to use in IHC-P and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting to hear back from you regarding your new results and whether they have improved or not..

If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Thank you for contacting us.

As we discussed over the phone, your protocol seems to be fine.

You had indicated that you would like to contact the manufacturer of the secondary antibody as well as try one more time. Maybe using less secondary and primary antibody could help, as well as increasing the blocking time.

Please let me know what additional information you might find out as well as if the result might improve after the next try.

As I had mentioned over the phone, I'd be happy to offer you either a free of charge replacement, a credit or a refund if the results remain unsatisfactory.

I wish you good luck and look forward to hear back from you.

Merci pour votre requête.
La méthode de démasquage est une étape très importante à optimiser. Nous conseillons donc en générale d'optimiser cette étape dans chaque laboratoire.
Effectivement, si un anticorps avait fonctionné bien en HIER avec Citrate, les chances sont qu'il fonctionne aussi pour vous en Citrate. Ils se peux qu'il ne fonctionne pas en EDTA, mais ceci reste à tester. Pour conclure, je peux donc conseiller de tester les tampons que vous avez à disposition. Par contre pour faire la mise au point de cet anticorps dans votre laboratoire, je vous conseille de tester les différents tampons en parallèle.
Pour illustrer ceci, je peux vous recommander de regarder la publication ci-dessous. Les chercheurs ont comparé les différent tampons pour différents anticorps.
Emoto et al., J Histochem Cytochem. 2005 Nov;53(11):1311-21.
Nous vous encourageons aussi à soumettre une Abreview par le biais de notre lien internet suivant, pour partager aussi votre expérience avec ces anticorps :
https://www.abcam.com/abreviews
Nous espérons que ces informations aideront. N'hésitez pas à me recontacter pour plus de renseignements.

Merci de nous avoir contactés.
Pour le démasquage antigène nous recommandons la méthode HIER avec du tampon Tris-EDTA pH 9.0. Comme le montre une publication de 2005 (Emoto et al., J Histochem Cytochem. 2005 Nov; 53 (11) : 1311-21, tableau nº2) le tampon Tris-EDTA pH 9.0 est la méthode qui est la plus "universelle" pour obtenir un démasquage correct des antigènes en IHC-P.
Concernant les séquences immunogènes, malheureusement les séquences exactes sont confidentielles mais nous pouvons vous informer que le peptide utilisé pour produire l'anticorps ab15098 est dans la région aa 150-211 de la protéine Cluadin 1 référence UniProt O95832. Le peptide utilisé pour produire ab23355 dérive de la région aa 96-150 de la Claudin 23 humaine, référence UniProt Q96B33.
Les images des Western Blot obtenus avec ces anticorps sur échantillons humains sont sur les fiches techniques respectives : www.abcam.com/ab15098 et www.abcam.com/ab23355 .
J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

Thank you for getting back to me. Generally if an immunogen forms an alignment of 6 or more consecutive residues I would consider it possible that the antibody would react against the homolog of the species. However, if it is a polyclonal antisera this is not always the case as given its polyclonal nature it is a mixture of immunoglobulin molecules. I would be very interested to know the results of your sequence comparisons as I could not find an immunogen that carried significant sequence similarity to warrant the suggestion of the application of the antibody to fish. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you for your enquiry. I have performed a sequence analysis of your fish sequence with all of immunogens used to raise our antibodies against Claudin. Unfortunately none had significant sequence similarity that would make me consider reactivity likely. I am sorry that I cannot be of more assistance on this occasion.

Thank you for your enquiry. The antibody originates outside of Abcam and the originator of the antibody does not wish to make the amino acid sequence available as it is considered commercially sensitive information. I can tell you, however, that the identity between the immunogen and bovine sequence is 17/19 (89%) for ab15106. The homology is similar for ab15098. I would predict that the antibodies may recognize the bovine sequence as this homology is quite high, however, I cannot guarantee this as it is possible to have such high homology and little cross-reactivity. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.