Asynchronous HeLa were paraformaldehyde fixed, permeablized, immunofluorescently labeled and counterstained with DAPI. No nuclear staining signal was apparent indicating that the antibody does not appear to recognize its epitope under these conditions in these cells. Methanol-fixed samples were also tried and did not produce a nuclear signal.
Thank you for your feedback. We have also found that this antibody does not seem to be suitable for use in ICC/IF experiments (see application notes section). We hope this information might be useful for future users.