Anti-Nucleophosmin 抗体 (ab15440)

Thank you for your email.

The answer you have provided to customer is very well structured. Perhaps you could also tell customer, that the information or data Abcam has about these products is already on the datasheet, they do not hold any more data than this.

We also haven't tested these antibodies in brain tissue sections infact it is not feasible for us to test every antibody in each body tissue however as the target is expressed in brain so the antibodies are fully guaranteed to work

Please forward the following link to customer (http://www.proteinatlas.org/ENSG00000181163/normal). It shows the target is expressed in brain.

I hope this information will be helpful.

Thank you for your enquiry. You can do the experiment that you are describing but the primary antibodies must be of different isotypes. I have seen this experiment work best when you have one mouse antibody that is IgG and another that is IgM, and then you use an anti-IgG secondary and then an anti-IgM secondary. I do not believe there are any secondary antibodies that are specific for IgG1 versus IgG2, so you would have to do IgG versus IgM. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

I'm sorry to hear you are having a problem with ab15440. Thank you for your patience in waiting for my reply. It is difficult for me to determine at this point if the lack of staining is due to the primary or secondary antibody, or if the protocol could be adjusted to have success with the antibody. We have not had any complaints about the secondary or primary so it is difficult to determine. Since ab15440 is working in Western blot I am wondering if either the protocol needs to be adjusted or if the secondary is causing a problem for you. Immunofluorescence was added as an application because of the following review that a customer submitted: Sample Mouse Cell (embryonic stem cells) Application Immunocytochemistry/Immunofluorescence Blocking step: Serum as blocking agent for 2 hour(s) 0 minute(s) · Concentration: 10% Fixation: Paraformaldehyde Antigen retrieval step: None Incubation time: 12 hour(s) 0 minute(s) Dilution 1/100 Rating Good - I would definitely use it again Secondary antibody Name: Non-Abcam one was used: Alexa 568 Conjugation: do not use Dilution: 1/1000 The only major differences are that the review customer fixed the cells in paraformaldehyde, the primary incubation was overnight at 4 degrees, and the secondary was different. I would like to suggest the following modifications to your protocol: 1) Please use paraformaldehyde as a fixative instead of methanol:acetone as antibodies can be sensitive to the fixation time and method, 2) More importantly, please incubate in primary overnight at 4 degrees if you have not already done so, 3) Please confirm whether you have used the secondary with another primary to know that the secondary is giving you good results. Alternatively, please try a different secondary in IF so we will know if the secondary is causing the problems for you. Please let me know if this helps and do not hesitate to contact us for further advice.

Thank you for your enquiry. Our recommended positive control for our Nucleophosmin antibody (ab15440) is a HeLa or mouse fibroblast extract. I would recommend our HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7898). I hope this information helps, please do not hesitate to contact me should you require further assistance.