Question (32002) | Anti-N Cadherin antibody (ab18203)

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Question

Dear technical team, Our customer purchased this antibody and tested FACS. He just did blocking, antibody incubation and washing. https://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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