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 500μL 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 400μL 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)