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Anti-N Cadherin antibody (ab18203)

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2 questions for ab18203

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Question 1

Thursday 26-January-2012

I have bought your Abcam (ab18203) rabbit policlonal anti-cadherin antibody and used based on the information on your site "Predicted to work with cow". I have used it on my bovine samples and it works fine, at a 1/50 dilution. However, my non-selective IgG negative control also staining green! Could you please specify if it was tested in bovine or if any publications has done it and which negative control was used? Best wishes.

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; http://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.

Question 2

Tuesday 03-January-2012

Dear technical team, Our customer purchased this antibody and tested FACS. He just did blocking, antibody incubation and washing. http://www.abcam.com/N-Cadherin-antibody-ab18203.html He got good results but, he realized this antibody recognizes cytosol site by immunogen information. "Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human N Cadherin." He thought this antibody recognizes membrane protein but the loacation of this residues is cytosol by his information. So he asked that hedidn't treat anything(permeabilization or something)but how this antibody showed result as he expected. Please let me know. Best regards,

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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