Anti-Fibrillarin antibody - Nucleolar Marker (ab5821)

I appreciate your response, but that did not answer my question. I asked for downloadable copies of the data sheets, specifically because your webpage was giving me printing issues.

Thanks,

ANSWER:

Thank you for your response.

Unfortunately, we do not have the pdf format of our on-line product datasheets yet but IT Department will consider implementing it.

I have saved the on-line documents as word documents and attached the files to my response.

I hope this helps and if I can assist further, please do not hesitate to contact me.

Dear Sir or Madam,

I recently purchased some antibodies from your company, but I am now unable to locate the data sheets to keep for our records.

Is there a way to download them from the product page online? (ab95958, ab30310, ab95315, ab5821)

Thanks,

ANSWER:

Thank you for your enquiry and your interest.

The datasheets of these products can be found at our website:

http://www.abcam.com/fibrillarin-antibody-nucleolar-marker-ab5821.html

http://www.abcam.com/nop5p-nop56p-antibody-37c12-ab30310.html

http://www.abcam.com/nop58-antibody-ab95315.html

http://www.abcam.com/nhp2l1-antibody-ab95958.html

If you need any further assistance in the future, please do not hesitate to contact me.

Thanks a lot for your prompt reply. I actually used three different dilutions: 1:250, 1:500, and 1:1000 but none gave me nucleolus staining. I can only see diffused dots thoughout the cytoplasm and nucleoplasm.

  I really appreciate if you kindly give me a nucleolus marker antibody replacement. Either Nucleolin antibody (ab22758) or mouse fibrillarin antibody 38F3 should be great, as long as they are certified in immunofluorescence.

  Again, thank you very much for your generous help!  

ANSWER:

As requested, I have issued a free of charge replacement, ab4566, mouse fibrillarin antibody 38F3.

To check the status of the order please contact our Customer Service team and reference this number. You should expect the antibody on Tuesday of next week.

There is a specific cell fixation protocol for staining cells with this antibody. Your protocol (fixation with PFA for 30 minutes followed by permeabilization) may also work, but this was the protocol used to produce the three large images at the bottom of the online datasheet. In brief, cells are fixed with formalin at room temperature for only one minute, then further fixed with cold methanol for only one minute. The protocol is at the following link:

http://www.abcam.com/assets/popups/popup_datasheet_protocols_printer_friendly.cfm?pid=179&intAbId=4566

The online datasheet is here:

Click here (or use the following: http://www.abcam.com/index.html?datasheet=4566).

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

ab5821 (Fibrillarin antibody - Nucleolar Marker)

Is this product LPS-free?

ANSWER:

Thank you for your enquiry.

ab5821 has not been tested for the presence of endotoxins therefore I can not guarantee that it is LPS-free.

Please do not hesitate to contact us again if you have further questions.

Hello Bill, As I discussed with you few minutes ago I have a problem with Fibrillarin antibody (ab5221). Instead of regular staining of nucleolus I have a label around it. I have tried different fixation protocols (either 2% or 4% PFA, 70% Ethanol, 100% Methanol at -20 following with acetone, Methanol-Acetic Acid Mixture) all the time I am getting similar weird pattern, which is very different from what is shown on Abcam website. You can see it yourself at the attached image. Because of this problem I would like to replace this antibody with another aliquot. I may use any another type of fibrillarin antibody ( on your site are also listed mouse monoclonal antibodies ab4566 and ab18380). The reason why I ordered specifically ab 5221 antibody is because it was the only one which was illustrated by a high magnification image. On the other hand I can use any other antibody if you have better references on them.

ANSWER:

Thank you for your email. Would you be able to fill out the form at the link below so I can see exactly what protocol you follow?

http://www.abcam.com/index.html?section=ihc&pageconfig=technical&intAbID=5821&mode=questionaire

I agree that your staining is not correct but I am wondering if you followed the exact protocol we recommend. We recommend the following protocol: fix in PBS buffered 4% PFA for 5 minutes, permeabilize with 0.5% triton-PBS for 5 minutes, block with 5% milk / 0.2% tween for one hour. Primary antibody incubation at 1/100 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Can you please fill out the form above? We use this protocol and have had many customers successfully use it as well. Can you please try this protocol if you have not tried it already?

I look forward to your reply,