Question (30975) | Anti-STIL/SIL antibody (ab89314)

Go to datasheet (ab89314)

Question

We had ordered Rabbit anti STIL antibody 20ug; item code: ab89314 on 28 September 2011. The purchase order number for the same is*******. We have a complaint regarding the antibody. the exact details of the complaint are given in the attachments.

Answer

Thank you for contacting Abcam. I am sorry that you are having issues with ab89314 in western blot. Having looked over the data and protocol information you kindly sent, I have a few suggestions that may help: 1 - Although the predicted mol weight is 143kDa, the western blot data on our website shows that when the antibody is used against HeLa cell extract, it recognizes a band at 170kDa. From the image you provided there does appear to be a (faint) band at about 170kDa, which may be correct. 2 - There are a lot of non-specific bands in your image; one reason for this could be breakdown of your protein, as I noticed you were not used protease inhibitors in your lysis buffer. I would advise adding protease inhibitors to your buffer. 3 - An alternative explanation for the multiply bands, could be that the antibody is binding non-specifically and therefore I would recommend switching to a 5% BSA block and antibody incubation buffer. 4 - I would also recommend loading more of your HeLa cell sample into your gel. Currently you are loading 3ul od samples, I suggest increasing this to 10ul, which will increase the signal from the 170kDa band. I hope that these protocol tips prove to be helpful. If not under our Abpromise, this antibody is guaranteed to work in western blot on human tissue. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund. If there is anything else I can help you with, plea

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