Anti-alpha Tubulin antibody (ab15246)

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?

ANSWER:

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.

Hi, I’d appreciate some help searching your products page on your website!  I’m trying to find primary antibodies against the C-terminal ends of alpha and beta tubulin for immunogold electron microscopy, but your search engine gives me 180 results for anti-tubulin antibodies but no means of sorting them with respect to their epitope.  I’d rather not have to open each webpage for all 180 hits to find the antibody I need, is there an easier way?

This work will be done in mouse, rat and human tissues, so I need the primaries not raised from these species.

Many thanks,

ANSWER:

Thank you once again for your enquiry and for your patience.

I am sorry to confirm that for Beta tubulin, the antibodies we have available tested and guaranteed in EM and mouse, rat and human tissues are mouse monoclonals. I can suggest to consider trying the following rabbit monoclonal antibody which is tested in mouse, however I am sorry this is not tested and guaranteed for EM.

ab52623 Rabbit mono Anti-beta Tubulin antibody [EP1569Y]

Applications

Flow Cyt, ICC, IHC-P, IP, WB

Reacts with: Human, Mouse, Rat

Immunogen propriatory, but it is right on the C-terminus and about 15 residues long.

http://www.abcam.com/index.html?datasheet=52623 http://www.abcam.com/index.html?datasheet=52623.

For alpha tubulin, we have the following antibody tested in EM and mouse, rat and human but rasied in mouse. I am sorry the epitope has not been mapped:

ab80779 Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free

Mouse monoclonal

Applications: EM, Flow Cyt, ICC/IF, IHC-P, IP, WB

Reacts with: Human, Mouse, Rat, Chicken, Cow, Gerbil, Guinea pig, Pig, Xenopus laevis

http://www.abcam.com/index.html?datasheet=80779 http://www.abcam.com/index.html?datasheet=80779.

I understand you concerns that mouse on mouse can sometimes give non specific binding. However, this has been tested succesfully in mouse, and we are able to provide our guarantee for the results from mouse and EM.

Otherwise, it could be preferable to try an antibody raised in a different species. The following rabbit polyclonal is tested in mouse, rat and human and is raised against a peptide from the C terminus. However, I am sorry it is not tested and guaranteed in EM:

ab15246 Anti-alpha Tubulin antibody

Rabbit polyclonal

Applications: ICC/IF, IHC-Fr, IHC-P, IP, WB

Reacts with: Fruit fly (Drosophila melanogaster), Human, Mouse, Pig, Rat, Zebrafish

Immunogen: ALEKDYEEVGVDSVEGEGEEEGEEY

C terminal amino acids 426-450 of Human alpha Tubulin

http://www.abcam.com/index.html?datasheet=15246 http://www.abcam.com/index.html?datasheet=15246.

With regards to the search facility on the website, I understand it is not possible to search by immunogen or region. This is partly because although we produce many antibodies in house and can provide the immunogen sequence, we do source many externally and sometimes the immunogen information is propriatory (commercially sensitive). We can usually provide this information on individual customer request.

Thank you for your understanding. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Dear Technical Team,   Hope this e-mail finds you well!   One of our customer is working on the separation of the nuclear and cytoplasmic fractions from mammalian cells like HEK 293 (HUMAN) and PS Cells(Porcine-Pig) & wants to make sure that there is no contamination of nuclear fraction in cytoplasmic fraction and vice versa. For that she wants to probe nucleus and cytoplam specific proteins in western blotting.   Kindly advise the suitable antibodies which can detect the cytoplasmic proteins & not the nuclear.   Looking forward towards your valued response on it.   With Best Regards,  

ANSWER:

Thank you for your enquiry.

I can confirm that the loading control protein alpha tubulin should only be located in the cytoplasm. We have several antibodies for detection of alpha tubulin which have been tested in WB, human and pig. I can recommend to review the datasheets for further information.

ab15246 Anti-alpha Tubulin antibody Rabbit polyclonal Applications: ICC/IF, IHC-Fr, IHC-P, IP, WB Reacts with: Fruit fly (Drosophila melanogaster), Human, Mouse, Pig, Rat, Zebrafish Click here (or use the following: http://www.abcam.com/index.html?datasheet=15246).

ab7750 Anti-alpha Tubulin antibody [TU-01] Mouse monoclonal Applications: ELISA, Flow Cyt, ICC, IF, IHC-P, IP, WB Reacts with: Human, Mouse, Arabidopsis thaliana, Cow, Dog, Pig Click here (or use the following: http://www.abcam.com/index.html?datasheet=7750). ab80779 Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free Mouse monoclonal Applications: EM, Flow Cyt, ICC/IF, IHC-P, IP, WB Reacts with: Human, Mouse, Rat, Chicken, Cow, Gerbil, Guinea pig, Pig, Xenopus laevis Click here (or use the following: http://www.abcam.com/index.html?datasheet=80779).

If you require any further assistance to find a suitable nuclear marker, please do not hesitate to let me know.

I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Thanks you for your email and your offer of 30% discount. We would be interested in testing your rabbit anti alpha tubulin antibody code ab15246.

ANSWER:

Thank you for your reply.

As my colleague Dr Sarah Mardle has explained to you it is not our policy to provide small samples for testing. However, if an antibody does not perform as specified on the datasheet, we will provide a full refund or offer you a replacement of equivalent value.

We might consider to offer a discount if you could send us a brief description of your research project.

We look forward to hearing from you soon.

Thanks you for your email and your offer of 30% discount. We would be interested in testing your rabbit anti alpha tubulin antibody code ab15246.

ANSWER:

Thank you for your reply.

As my colleague Dr Sarah Mardle has explained to you it is not our policy to provide small samples for testing. However, if an antibody does not perform as specified on the datasheet, we will provide a full refund or offer you a replacement of equivalent value.

We might consider to offer a discount if you could send us a brief description of your research project.

We look forward to hearing from you soon.