Hello, I am trying to perform some IPs using Dynabeads G from invitrogen on Ago2 protein from Jurkat cells, and I need a negative control.
I have some rabbit Abs for my target protein (Ago2), and the question is do I also need a RABBIT total IgG as a negative control? Would IgG from other animals work for that purpose? Like goat, mouse or rat (looks like we have these here)
Thank you!
Hello
To make sure of your result , would prefer to use extra positive and a negative control, also this will help you to publication in journals with high impact factor
For the question about the use ( IgG) from others animals , yes Because the vast majority of primary antibodies are produced in just a few host animal species and most are of the IgG class, it is economical to produce and supply ready-to-use secondary antibodies for many methods and detection systems. From a relatively small number of secondary antibodies, many options are available for purity level, specificity and label type for a given application. But you must make sure the his accompanying catalog .
visit this link to read more information about this Issue
Good Luck
http://www.pierce-antibodies.com/products/secondary-antibodies/
yes, you should run the same IP protocol using normal rabbit IgG and run it on your western blot. If your IP was specific with the antibody that you used, you should not see the band in the normal rabbit IgG sample. If you see the same band in your IP samples of interest and in the normal IgG sample, you are pulling down non specific bands. You can purchase normal rabbit IgG from multiple companies.
Hello Mark.
You definitely need a Rabbit IgG as a Negative control, Try to get that from Invitrogen.
Good luck
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