Anti-GFP antibody [GFP01 (B34)] (ab3277)

Hi

I am looking for antibodies that can distinguish between the N-terminus

of YFP (GFP) the first 155 aa and the C-terminus aa156-against the first

156-240. These would be for the prominent Spli-YFP or Bimolecular

Fluorescence complementation (BiFC). Do you know whether you have a pair

of GFP antibodies raised against specific immungens within these regions?

Cheers

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ANSWER:

Thank you for your inquiry and please excuse the delay of my answer.

Most of our GFP (YFP) antibodies were generated using the full-length protein and I was checking if the epitopes of the monoclonal antibodies (ab291, ab3277, ab38689) were mapped.

Unfortunately, no epitope mapping was performed and I am only able to suggest one antibody that might be suitable for your purposes.

ab32146 is specific for the N-terminal part 1 - 155aa.

http://www.abcam.com/index.html?datasheet=32146 http://www.abcam.com/index.html?datasheet=32146.

Unfortunately, we do not have antibodies where I can guarantee that they are specific for the other regions of the protein.

I am sorry I could not provide a completely positive answer to your questions on this occasion and hope this information is nevertheless helpful.

Please let me know if you need any more information about ab32146 or any help purchasing it.

Dear Tech support,

One of our customers needs an anti GFP antibody that recognizes the c-terminal end only, I assume it's more likely to find suitable monoclonal antibody. The required applications are WB and ICC/IF

Is one of the following antibodies suitable:

ab1218

ab291

ab42560

Alternatively, please suggest adequate antibodies.

Thank you for your kind help.

Regards,

ANSWER:

Thank you once again for your enquiry.

I have checked all the monoclonal antibodies to GFP tested in western blot that we have available.

I am sorry to confirm that they are all raised against full length protein, and the epitopes have regrettably not been mapped. We would therefore not be able to guarantee these will be detecting the C terminus of the GFP protein.

I am awaiting a response from the originator for one more antibody. Should this provide any more helpful information, I will contact you and let you know.

I am sorry there are no suitably epitope mapped antibodies to confirm your requirements on this occasion. Should you have any further questions, please do not hesitate to contact us.

Unfortunately, I must have thrown away the films I developed from the membranes I developed. In an attempt to get some usable data from the blot I used another anti-body to see what was happening.

I will repeat my experiment- with positive and negative controls with cells expressing a GFP fusion protein and a cell of the same type not expressing the fusion protein. I will try using the antibody at a number of dilutions - 1:100, 1:500, 1:1000, 1:2000, and 1:3000. I will also include a control for the secondary without antibody bound. I looked over your protocol recommendations- I will give them a try. Who knows maybe, the antibody will work. I unfortunately, won't have time for the next couple of days to optimize the protocol, I am trying to make a deadline. I will write back as soon as I have the results.

Thanks for writing back so quickly and thanks for helping me with my concerns.

ANSWER:

Thank you for your email. If you continue to experience problems with this antibody, please do let me know.