Alkaline Phosphatase Staining Kit

Alkaline Phosphatase Staining Kit is designed for the rapid and sensitive detection of osteoblasts differentiated in culture from bone marrow cells. Bone forming osteoblasts and bone-degrading osteoclasts can be distinguished by the expression, respectively, of alkaline phosphatase and tartrate-resistant acid phosphatase (TRAP). CosmoBio’ s TRAP staining kit (PMC-AK04F-COS) is ideal for rapid and convenient detection of the latter.

Simple staining protocol

1. Remove culture medium. Wash each well 3 times with 1mL PBS.
   


2. Add 500μ L of 10% formalin neutral buffer solution to each well and fix for 20 minutes at room temperature.
   


3. Remove 10% formalin neutral buffer solution and wash each well 3 times with 2mL deionized water.
   


4. Dissolve 1 vial of Chromogenic Substrate in 5mL Substrate-containing Buffer.
   


5. Add 400μ L Chromogenic Substrate to each well.
   


6. Incubate at 37° C for 5-20 minutes. Adjust incubation time until stained.
   


7. Wash with deionized water to stop the reaction.
   

References

1. A. S. H. De Jong, M. Van Kessel-Van Vark and A. K. Raap. Sensitivity of various visualization methods for peroxidase and alkaline phosphatase activity in immunoenzyme histochemistry. The Histochemical Journal Volume 17, 1119-1130 (1985)