The TRAP Staining Kit is used for the staining of Tartrate-Resistant Acid Phosphatase in osteoclasts. Bone mass is controlled by the balance between the activity of osteoblasts bone formation and the activity of osteoclasts.
Alkaline phosphatase (ALP staining kit, Cat.No.PMC-AK20-COS) and tartrate-resistant acid phosphate are used as markers for osteoblasts and osteoclasts, respectively
Figure 1: TRAP staining of Osteoclast
|Tartrate-containing Buffer||50 mL||4C (Do not freeze)|
|Chromogenic Substrate||10 vials||4C|
One kit contains regents for 10 × 96-well plates
#Additional Materials Required
・10% Formalin Neutral Buffer Solution
・Phosphate Buffered Saline (PBS)
・Distilled or deionized water
1. Remove culture medium. Wash each well once with 100uL of PBS.
2. Add 50uL of the 10% Formalin Neutral Buffer Solution to each well and fix for 5 minutes at room temperature.
3. Wash each well with 250uL of deionized water. (× 3 times)
4. Dissolve 1 vial of Chromogenic Substrate with 5mL of Tartrate-containing Buffer.
5. Add 50uL of Chromogenic Substrates to each well.
6. Incubate at 37℃ for 20-60 minutes. Adjust incubation time until stained TRAP is clearly showing the result in figure 1.
7. Wash with deionized water to stop the reaction.
Note: Excess incubation will be cause of over stained.
TRAP Staining Kit