Anti-Ago2 / eIF2C2 antibody (ab5072)

I acquired the Ago2/eIF2C2 - (ab5072) antibody and I have some doubts about it.

1. When I chose the antibody, the specie reactivity was Drosophila AND Human (I received a datasheet with this information....). However, I looked today and the new datasheet says: "Reacts with Fruit fly (Drosophila melanogaster). Does not react with Human.". What happened? I intended to use this antibody with human cells!

2. Usually, EIF2C2 is viewed in the citoplasm, associated to processing bodies (PMID:16880270 is cited in your datasheet as a reference where ab5072 was used... and it was co-localized in the citoplasm with GW182). But in ABCAM datasheet, it cellular localization is: "Nuclear". So... i think that I am missing something.... I would like to know how Schneider et al. were able to use ab5072 and see something in the cytoplasm, because I am using the same antibody in human cells (and it reacts with some protein... now I am not sure that it is really Ago2/EIF2C2), but I just see signal inside the nucleus.

I hope to count with you to solve all this mistery...Thanks for your kind attention,

ANSWER:

Thank you for your enquiry.

I sincerely apologize for the error. We just received information regarding the immunogen sequence for this antibody. On investigation by our laboratory, it seems that it does not have any alignment to the human protein. Therefore, this antibody is not expected to react with human samples and therefore, the datasheet has been updated to show the changes.

We also received a review from a customer who managed to obtain results with human samples but after confirming the sequence alignment, we deduced that the results obtain by the customer was probably due to non-specific binding.

I am unsure about the findings of Schneider MD et al (2006) but our information was taken from SwissProt database. If you have any questions regarding the findings from the authors, I would suggest that you contact them for further details.

I am truly sorry that we had a mistake on our datasheet and I am more than happy to offer you a free replacement product of your choice that has the same value as this ab5072, or a refund. Please let me know which method you would prefer and please confirm your purchase details (purchase order number, shipping address/purchasing agent), so that I can immediately issue you the replacement or refund.

I look forward to hearing from you again in order to resolve the matter.

I am using ab5072 (Rabbit polyclonal to Ago2/eIF2C2) for some assays and wanted to order a good control antibody from you. It would have to be an affinity purified Rabbit polyclonal antibody, preferably (though I guess not necessarily) against a Drosophila target. Since it is a control, it should not target any components of the RNAi machinery. Something like a transcription factor would be ideal since I am looking at protein expression at a time when Drosophila are transcriptionally silent. I was hoping you could suggest some candidates from your catalogue. The concentration would have to be at least 0.5 mg/mL.

Thank you!

ANSWER:

Thank you for your enquiry.

Assuming that you are interested in an antibody to control for non-specific binding I would like to recommend that you use an affinity purified rabbit polyclonal antibody against an irrelevant target such as one of the epitope tags. These are obviously foreign to Drosophila and will serve to highlight any non-specificity in your experiments.

For example: V5 tag antibody - ChIP Grade (ab15828)

This is an immunogen affinity purified sera and supplied as a range: 0.20 - 0.50 mg/ml

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.