Improving method of immunoprecipitation
Immunoprecipitation is a commonly used technique for studying proteins, and it is a method for purifying and enriching the protein of interest using antibody-specific reactions. After the antibody binds to the corresponding protein in the cell lysate or the expressed supernatant, it is then incubated with protein A / G (ProteinA / G) or secondary antibody-coupled agaose or Sepharose beads, and the beads-protein A / G or two are obtained by centrifugation After washing the anti-antibody-target protein complex, the pellet is resuspended in the electrophoresis sample buffer and boiled for 5-10 minutes. Under the action of high temperature and reducing agent, the antigen and antibody are dissociated, and the supernatant is collected by centrifugation. This includes antibodies, proteins of interest and a small amount of contaminated proteins.
If the protein sample is not pre-treated with beads before incubating the antibody, it will cause some non-specific proteins to bind to ProteinA / G, thereby interfering with the test results. In addition, if antibodies and beads are added at the same time, sometimes the precipitation efficiency is not high.
Method improvements:
1. Pretreatment: Before adding antibodies and beads, add 20-30 microliters of 50% concentration of bead suspension in protein lysate and incubate for one hour on a 4 degree shaker. For laboratories that often do IP or CO-IP, the beads used for the first pretreatment of the same sample can be collected and stored at four degrees for pretreatment when the same protein lysate is precipitated next time.
2. Incubate the antibody and beads separately. First add the appropriate amount of antibody to the protein lysate and incubate overnight on a 4 degree shaker. Then add the right amount of beads and incubate for 2 to 3 hours on a 4 degree shaker.
The table placemat is a kind of thing that can protect, decorate table , it makes of PP material.Placemat has stronger frictional force to prevent glass, porcelain cup to slide, it can also protect desktop not to suffer very hot bad.The placemat can be used in various occasions, such as home ,hotel,restaurant,wedding party place and so on .
Table Placemats,Home Simple Table Mat,High Grade Color Printing Pad,Kitchen Waterproof Grease Pad
shaoxing chaofeng stationery manufacturing CO.,LTD. , https://www.chaofengstationery.com