Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

The most recent vial of ab7291 gives no signal in blots of C2C12 lysates.

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I have issued a free of charge replacement with the order number *****.

To check the status of the order please contact our Customer Service team and reference this number.

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.

Is this tubulin antibody likely to cross-react with C. elegans?

ANSWER:

Thank you for contacting us.

I have not been able to find the C. elegans alpha tublin sequence to align with the epitope this clone recognizes (located within the range of chicken alpha tubulin specified on the datasheet). However, I did find a reference that is listed on the datasheet of another version of this clone, DM1A, with the catalogue number ab64503, a FITC-conjugate.

Click here (or use the following: http://www.abcam.com/index.html?datasheet=64503).

The paper is:

Dumont J et al. A kinetochore-independent mechanism drives anaphase chromosome separation during acentrosomal meiosis. Nat Cell Biol 12:894-901 (2010). PubMed: 20729837

The authors used the antibody for immunofluorescent microscopy. Until we receive more data, we will not guarantee ab7291 for reactivity with C. elegans, but I think this paper is encouraging. If you do decide to try it, please consider sending us a review of your resutls. For instructions on how to do this, please see the page at the following link:

www.abcam.com/abreviews

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

New vial shows no bands in WB.

ANSWER:

Thanks for your call today and for letting us know about the problem with the most recent vial of this antibody.

As we discussed, I am sending a vial from a new lot on the order ***, which should arrive promptly.

Please keep me updated about how this new vial works, and let me know if there is anything else that we can do for you.

Phone call regarding the protocol used to obtaining the staining with HeLa cells with ab7291.

ANSWER:

Thank you for contacting us. As requested the protocol currently used to stain the HeLa cells with Anti-alpha Tubulin antibody [DM1A] (ab7291) is as follows:

The cells were fixed in 100% methanol for 5 minutes and then washed with PBS. The cells were then permeabilised with PBST (0.1%) and blocked for 1 hour in 1% BSA/10% normal serum/0.3M glycine. The block was then washed off using PBST. The primary antibody (5µg/ml) diluted in 1 BSA/PBST was then incubated with the cells overnight at 4 degrees C. The secondary antibody was diluted in the same buffer as the primary antibody and incubated for 1 hour at room temperature.

This is not exactly the protocol used to produce the image displayed on the datasheet but one that has been further optimised since and is used fro quality control of the antibody. I have attached an image obtained using the protocol described.

I hope this information has been of help. If you are having any difficulty obtaining the staining expected using this antibody or you'd like any further information please do let us know.

Hello, Recently we ordered a new aliquot of Ms mAb alpha tubulin catalog number ab7291 lot number GR16134-1.  We have previously had good results with this antibody but this new aliquot has not been consistent and works with a much lower efficiency.   Is there a way for us to get another different lot of this antibody?  We would be willing to order another tube but I want to make sure that we are not going to be shipped the same lot with potentially the same problems.   Thank you for your assistance.

ANSWER:

Thank you for contacting Abcam. I am sorry that the last lot did not perform aswell as previously. I can confirm that we have a different batch of ab7291 in stock and that if you were to order a new vial of the antibody then you would not be getting the same lot as you previously received. If there is anything else I can help with, then please let me know.