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Question (62888) | Anti-mTOR antibody (ab2732)

Dear Abcam technical support

I'm interested to buy an antibody against mTOR which works well in Western blot.

According to NCBI the molecular weight of mTOR is 289kDa.
The antibodies against mTOR you are selling i.e. ab2732 show specificity against a protein of 255kDa.

I was wondering where this difference is coming from?

And which of the antibodies against mTOR would you advice me to buy for use in Western blot?

Many thanks

Thank you for contacting us and your interest in our products.

When trying to detect a protein of the size of mTOR in Western blotting, there is some movement of the band that would be expected away from the predicted band size. Due to how the ladder and protein runs in the SDS-PAGE gel there can be quite large variations in proteins of this size so the size of 255 kDa observed with ab2732 would not be unexpected. The antibody has been used in the staining of HepG2 cells as well as rat testis, supporting it detecting the expected protein.

I have had a look through the antibodies we have against this target and I would suggest also taking a look at the mouse monoclonal ab87540 which has been used in Western blotting and produced a clean band of ˜268 kDa with HEK293 and human liver tissue (amongst other samples).

However, I'd like to reassure you that both the ab2732 and ab87540 would be covered by the Abpromise guarantee if you intend to use it in Western blotting with human samples. More information on this guarantee can be found from the following link:

http://www.abcam.com/abpromise

I'd also like to draw your attention to our protocol booklet which has some useful advice when performing Western blotting with such large proteins. This booklet can be accessed from the following link:

http://www.abcam.com/index.html?pageconfig=resource&rid=13111

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.