I need to load 120 ug of proteins in a protein gel. Do I need some precaution for the protein migration and western-blot?
Yes, you can load 120 ug of proteins in a protein gel. However, the mobility of your protein might be changed and distorted after that. Depending on your specific protein mixture, you might get good results or not so good. For example, if you have one highly dominating protein, such as albumin, you will get bad results. If all proteins in your mixture are approximately in the same abundance order you might get relatively good results. If your goal is just answer the question if your protein is present in your mixture or not, your can go ahead with 120 ug. You will get the answer. If your goal is to get some quantitative estimate about the relative abundance of your protein in the mixture, or its approximate MW, I would not recommend the load with 120 ug. Any presence of salts, detergents, DNA, polysaccharides, or cell membrane or sub cellular fragments will also distort the mobility of your protein. The load of 120 ug might also lead to some non-specific bands in your western blot (depending on the Ab used) because of some potential cross-reactivity of your Ab with other proteins in your mixture (because of their relative abundance).
The amount of protein that can be loaded in one lane of a gel also depends on the size of the gel, its thickness, and the width of the lane. The greater any one of these dimensions is, the more protein can be loaded.
Again, the volume which would go into the well for 120ug you have to see. Protean II XI large gel apparatus from BIORAD would solve the problem from my personal experience and also the proteins should be well-balanced ratio of different proteins and not some of dominating nature which may overshadow and reduce the resolution of lesser proteins.
Please explain what you mean by "proteins". And also the gel apparatus that you plan to use. And also the western detection system you plan to use.
I have loaded 200ug of protein in midi gel but try to load in less volume for which you should try 6X dye instead of 2X or 3X dye. U should also use midi/maxi gel as in mini gel, protein will run in a improper way due to which u will not get clear band (Just imagine so many participant are running for small race, due to which there will be so much confusion and distortion within participant)
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