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TRAP Staining Solution For Osteoclasts Staining kit

  • G1050-50T

Product Description

Principle

TRAP is one of the six acid phosphatase isozymes, the type VI. TRAP is rich in osteoclast, usually as an indicator to identify osteoclast. Osteoclast stained with TRAP staining method show positive claret-red in cytoplasm and negative in nucleus. 

Package content

Code

Name

Size

G1050-1

Acetic acid buffer solution

18 ml

G1050-2

Pararosaniline solution

500 ul

G1050-3

Sodium nitrite solution

500 ul

G1050-4

Naphthol AS-BI phosphate

20 mg

G1050-5

N,N-dimethyl formamide

1 ml

G1050-6

Potassium sodium tartrate

0.282 g

Preservation

Valid in 12 months when stored at 4℃ and kept in dark condition. 

Reagents should be prepared before use

1. Mix up a solution of equal parts of Sodium nitrite and Pararosaniline, marked as solution B.

2. Dissolve the Naphthol AS-BI phosphate in 1 ml of N,N-dimethyl formamide, marked as solution C.

3. Preparing incubate solution: mixing the reagents including Acetic acid buffer solution (18 ml), solution B (1 ml) and solution C (1 ml) well, adjust to pH=5.0 by NaOH/HCl (1 M). Add this obtained mixture solution into the tube containing potassium sodium tartrate (0.282 g), shake well and filtrate. Then obtained the TRAP incubation solution.  

 

Procedure

1、Deparaffinize and rehydrate to distilled water, wash for several minutes.

2、Incubate slides within the incubation solution at 37℃ for 50-90 min.

3、Wash in tap running water for 2-5 min.

4、Counterstain nucleus with hematoxylin solution for 1-2 min, differentiate in hydrochloric acid, and back to blue in ammonia solution.

5、Dehydrate with routine method, then clear and mount.

 

 

Results

The area of positive acid phosphatase activity is claret-red and nucleus is blue.

Notes

1、 This staining kit applies in bone tissues which has been decalcified by EDTA. Acid decalcifying solution have adverse effect on acid phosphatase activity of bone tissues, therefore is not suitable for this staining solution. 

2、 Slides must be washed with distilled water before incubating in staining solution. 

 

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