Alkaline Phosphatase Staining Kit - Cosmo Bio Co.,Ltd.

Antibodies

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Alkaline Phosphatase Staining Kit

Catalog No.: PMC-AK20-COS
Size: 1KIT
[12 well plate x 10]
Price: ¥37000
$494
catalog info: Catalog 2012-p142
Storage: 4C DNF
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Purpose: Detection of Osteoblast
component: Substrate-containing Buffer (50 ml) x 1 bottle
Chromogenic Substrate x 10 vials
One kit contains regents for 10 ~ 12-well plates
#Additional Materials Required
E10% Formalin Neutral Buffer Solution
EPhosphate Buffered Saline (PBS)
EDistilled or deionized water
Additional Materials Required
E10% Formalin Neutral Buffer Solution
EPhosphate Buffered Saline (PBS)
EDistilled or deionized water

Other:

 Background 
The Alkaline Phosphatase Staining Kit is suitable for the staining of alkaline phosphatase in osteoblasts. Bone mass is controlled by the balance between the activity of osteoblasts bone formation and the activityof osteoclasts.
Alkaline phosphatase and tartrate-resistant acid phosphate (TRAP: TRAP staining kit , Cat. No. PMC-AK04F-COS) are used as markers for osteoblasts and osteoclasts, respectively.
 Protocol 
(12-well plate format)
1. Remove culture medium. Wash each well three times with 1mL of PBS.
2. Add 500uL of the 10% Formalin Neutral Buffer Solution to each well and fix for 20 minutes at room temperature.
3. Remove 10% Formalin Neutral Buffer Solution. Wash each well with 2mL of deionized water. (~ 3 times)
4. Dissolve 1 vial of Chromogenic Substrate with 5mL of Substrate-containing Buffer.
5. Add 400uL of Chromogenic Substrates to each well.
6. Incubate at 37 for 5-20 minutes. Adjust incubation time until stained ALP 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.



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ALP staining of 3T3-E1

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Reference:

•  A. S. H. De Jonget al.,The Histochemical Journal Volume 17, 1119-1130 (1985)
 
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