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Could you, please, help me with making a right choice of anti-alpha tubulin antibody. I need to make double immunostainings in Drosophila S2 cells to visualize co-localisation of the protein of my interest with mitotic spindle in dividing cells. I found that anti-alpha-tubulin antibody is mostly used for visualizing microtubules. For my application I need anti-alpha tubulin antibody produced in rabbit. In Abcam catalog I found only one antibody ab15246 that is indicated to recognize Dm antigen. But the datasheet for this product is lacking indication on how to use this antibody for fluorescent immunostainings. The PubMed reference 17785506 provided contains no details on that question either. On the other hand datasheet for antibody ab18251 contains an Abreview where this antibody was successfully used for immunostaining Drosophila embryos as well as details on antibody dilution. But in the datasheet itself it's not indicated that ab18251 reacts with Dm. So which antibody I should order to bring my experiment to success?
Asked on Sep 21 2011
Thank you for contacting us. We would guarantee both ab15246 and ab18251 to stain cells from drosophila via immunocytochemistry (ICC/IF). I am not sure why drosophila wasn't added as a tested species to the datasheet for ab18251 after that Abreview was added, but this was an oversight and I have since updated the datasheet to include drosophila as a tested species. As for the references for ab1546, we know of them testing drosophila by WB and IHC-Fr, not ICC/IF, per se. But we would still guarantee the antibody to work in drosophila and ICC/IF since both the species and application were tested, although independently. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3048163/pdf/cm0068-0001.pdf http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2562427/pdf/nihms70131.pdf If you're looking for an ICC/IF protocol to use with ab1546, you can look at the below two articles, tested in mouse and human, and just apply that same protocol to your cells from drosophila. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2872425/pdf/pnas.200910941.pdf http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2851247/pdf/nihms120363.pdf Both of these antibodies would be covered under our Abpromise to use them with drosophila in ICC/IF so if you ran into any difficulty, we would be happy to replace or refund them for you within 6 months of your purchase. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Sep 21 2011