I want to know the basic differences between these techniques and their application in molecular biology and pharmacology?
Hi, I will try to answer you but it is not simple. Firstly, you should distinguish from ihc/if and ip; therefore in both the cases you use an antibody that recognise your antigen. In ihc/if you want to find where is you antigen. In this case you use a primary antibody that is specific for you ag and then you will recognize your primary ab through a secondary ab that could be conjugated with a fluorofore (in thiscase you are doing a if) or it could be bound to biotin (in this case you are doing ihc). in theis last case, you need avidin and then dab for seeing something. With if you could decide also to recognize more than one ag, i.e if you want to see if a particular receptor is inside a neuron, you will put primary ab for the antigen you like and an other one for neurons. You will aquire images using microscope. If is mostly use for having a qualitative idea of your antigen localization; on the contrary, ihc is used when you would like to have a quantitative idea because you can aquire images and quantifying your marker through specific programmes as imagej. As regarding ip, this method is useful if you would like to separate your antigen from a pool of proteinsl for example if you have plasma samples and you need to quantify albumin, you could decide to separate albumin from all the other proteins. In this case, you need protein a or protein g or a mix of both depending on what you need to separate (is the antigen an IgG?). These two proteins are needed to increase the strenght of the complex primary ab-antigen. After an ip you should detect your protein through a wb.
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