In Western blot analysis, protein identification is based on antibody and antigen reactions. Proteins are separated on polyacrylamide gels and are transferred (blotted) to a nylon membrane. The membrane is exposed to solutions containing primary antibody, followed by a secondary antibody coupled to an enzyme. The membrane is then soaked in a substrate solution to develop the color reaction, which results in identification on the antigen protein band. The molecular weights of the visible bands are measured using prestained protein markers of known molecular weight. This kit does not require an electrotransfer apparatus.
For 6 blots.
Time required: electrophoresis 60 minutes, blot overnight, detection 2.5 hours.
Kit includes: instructions, negative control, all samples & antibodies, various reagents and buffers, membrane and filter paper.
All You need: 3 polyacrylamide gels (12%), vertical gel electrophoresis apparatus, power supply, adjustable micropipet with fine tips, laboratory glassware, metric rulers, distilled or deionized water, glacial acetic acid, methanol.