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Dear Sir, I am using your antibody Akap12. With this mail i am attaching western blot figure which was developed using ab49849 antibody. I would like to confirm from you which is the exact Akap12 band. In your product details you have mentioned the molecular weight of Akap12 is approximately 300Kda (predicted is 191Kda). I have used high range molecular weight marker in one side and the other side is colour marker. In this figure which band is the exact Akap12 band (Whether first red arrow from the top or second arrow from the top). Figure 1 and 2 are same with different time exposure time. I have used 7% gel for this study. The sample used is cell lysate, not animal tissue. I will be happy if the reply is at the earliest.
Asked on Oct 20 2011
Thank you for contacting Abcam. I have looked at the western blot image that you attached and I believe that the Akap12 band is actually the large band a the top of the image, not marked by a red arrow. If you would like my opinion of which band is the closet to 191kDa, then I would chose the top red arrow, but again I believe the actual Akap12 band is higher. Based on the datasheet and the references listed on the datasheet (PubMed: 21831305), the band that is of interest is at 300kDa. To achieve an image that has better resolution, you could rerun the gel and this time let the gel run longer, so that you get rid of all the low molecular weight products and gain a greater separation of the higher molecular weight proteins. Also using a molecular weight marker, that goes to 300kDa would also allow more accurate sizing. Please let me know if there is anything else I can help you with.
Answered on Oct 20 2011