Corporate Headquarters
Vector Laboratories, Inc.
6737 Mowry Ave
Newark, CA 94560
United States
1) Cell monolayers are cultured in the 96-well microtiter plate. Each well can then be treated with stimulants, inhibitors, etc. The cells are then fixed and blocked. |
2) Antigen specific primary antibody is added and binds to the target antigen on the cells surface. |
3) The wells are washed and a HRP-conjugated secondary antibody is added which binds to the primary antibody. |
4) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow. |
1) Cell monolayers are cultured in the 96-well microtiter plate. Each well can then be treated with stimulants, inhibitors, etc. to induce expression of the phosphorylated target. The cells are then fixed and blocked. |
2) Phospho-specific or pan-specific primary antibody is added and binds to the target antigen on the cells surface. |
3) The wells are washed and a HRP-conjugated secondary antibody is added which binds to the primary antibody. |
4) The wells are washed and a solution containing TMB is added. The TMB reacts with the HRP changing from colorless to a blue color. An acid stop solution is then added, the reaction stops, and the blue color changes to yellow. |