Western Blot Analysis

The western blot analyze what proteins and how much of the protein are in a gel. A western blot is very sensitive to proteins with histidine or HIS tags. Anything that contains histidine will show up in the gel. The western blot is evidence to whether or not if IL-8 is in the gel. The importances of the western blot is to show how well the proteins reacts to a disease. The first step for a western blot is to run an unstained PAGE gel. Western blot uses gel electrophoresis or electrical to arrange the proteins in order by sizes and soak up the protein into the membrane which is referred to as blotting. When blotting is done the membrane is treated with an antibody(s) to only stick to the protein that is important (IL-8 and HIS Tag). After treating
This shows that IL-8 does exist in the sample. PAGE GEL The PAGE gel can be use in a western blot and also can be compared to the western blot. The difference between the western blot and gel is that the western blot focus on the protein IL-8 only and the gel has other proteins that shows up with IL-8. The PAGE gel contains undiluted induced (non-control), undiluted uninduced (control), reducing sample buffer (RSB), sodium dodecyl sulfate (SDS), kaleidoscope (ladder), and IL-8 (protein). Lanes: Proteins shows up in all ten of the lanes in the gel. Each lane add up to 40uL. Lanes one through six with undiluted induced bands are getting lighter as the proteins goes through lanes one through six. Lane seven is the lightest band of the undiluted uninduced lanes. The undiluted uninduced lanes have IL-8 and other proteins. Lane eight with undiluted uninduced is the darkest band out of the gel. Lane nine is the kaleidoscope which is a ladder with blood proteins and is a guild to show what proteins are in the gel. Lane ten has one band of IL-8 only.
32 ul undiluted induced, 8 reducing sample buffer
16 ul undiluted induced, 16 ul SDS, 8ul