Common Problems in Extraction of Protein Assay Kit |
Posted: August 4, 2016 |
Below, I listed some of the common problems and solutions of protein assay kits extraction, hoping to help your experiments. Q: how to grind tissue liquid nitrogen? A: tissue block is frozen in liquid nitrogen, then knock it into small pieces to place in the mortar and pour liquid nitrogen. Pour the liquid nitrogen and grind at the same time. Keep mortar with liquid nitrogen, or let tissue does not melt. Q: if membrane protein does not dissolve in loading buffer, or there is the formation of precipitation. How to do? A: degenerate for 1 hour with 60 degrees; if it does not work, the SDS of loading buffer can be replaced by LDS. Q: how many proteins extracted by cells are enough to make western blot? A: generally 5 x 106 is enough. But it depends on what kind of cells. Some cells are very small, and some cells are very large. Conventionally, 105 cells are needed on one pore of WB. But it also depends on the target protein and the way of extraction. If it is a membrane protein or a protein distributed in certain cells, the number should be increased. Q: can the same protein sample carry out two kinds of factor Western Blot detection at the same time? A: yes, even more than ten kinds of samples can be measured simultaneously. Q: how long can protein be stored after extraction? A: a year is OK. Two points: it can not be hydrolyzed by protease; it can not be contaminated by bacteria. Q: to do WB after extraction. What is the total amount of the protein? Is it related to the expression of target protein? Q: do I have any special attention when I make western of the tissue sample protein? A: it is necessary to grind and handle with homogenate so that protein solubility will be better. The protease activity in the tissue is strong, so we need to pay attention to the inhibition of protease activity. About the author: In order to speed up the process of research, Creative Biostructure provides high-quality protein kits, protein crystallization kits, membrane protein crystallization kits, etc...
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