I stained the cells with different VCAM-1 antibody, immediately after Ficoll or the day after. In both cases I found no signal.
Be sur ethat your cells express the relevant protein (a q PCR will give you an answer). If you are sure that you cells express it then probably you have a technical problem. Either you damage the VCAM1 protein by expossing you cells to culture enzymes for long period or you do not allow sufficient time to Moab to be atctached. Also be sure that the clone of your Moab is bond to your wanted VCAM1.
Thank you for the suggestions. I tried to follow your indications but I was not able to solve the problem. Could it be that the problem is that we are getting the bone marrow by post and so we are isolating the cells the day after? Maybe VCAM is already distrupted when we are doing the staining.
Hi Luca.
I hope you will also share your experience with the antibodies on www.pAbmAbs.com? Here, scientists can share information about the antibodies they use in their research. If these antibodies against VCAM-1 did not work you can save other scientists time and money by sharing this information. Your input is therefore very valuable for other scientist when they are looking for reliable antibodies.
As a bonus a can let you know that all reviews received before dec 15th will enter a competition where the winner will receive €500 for personal spending. Every review counts as a lottery ticket-also reviews on antibodies that have already been reviewed by other scientists before you. It is quite simple to review. All you have to do is go to the website, click "submit review" and here it is described how to design the review.
Kind regards,
Simon