4) Buffer for ACP 30 ml
0.5 M Sodium acetate, pH5.2
5) Buffer for ALP 30 ml
0.2 M Tris-HCl, pH9.5, 1 mM MgCl
2
* The concentration of Tris-HCl has changed for keeping pH stability from lot. 011.
6) Microplate (96-well) 1 plate
Used in sample dilution or container for reaction
The plate is reusable after soaking in 1 % sodium hypochlorite solution overnight.
Reagent required but not supplied in the kit
Stop Solution:0.9 N NaOH
*
Prior to starting the assay, a customer is required to prepare Stop Solution by himself.
*:This solution is corrosive. It may cause inflammation when it contacts with
skin. When it comes to contact with hands or mucous membrane, immedi-
ately wash away with large amount of water and follow instructions provided
by doctors.
IV. Storage
4 ℃
V. Preparation of Reagents
1) All reagents should be brought to room temperature before use.
2) Preparation of substrate solution
Dissolve 1 vial (24 mg) of [1] pNPP (
p-nitro-phenyl phosphate) substrate in 5 ml of
the enzyme buffer to be assayed ([4] or [5]) , and use this as the substrate solution
(substrate concentration:12.5 mM) . When used for tartrate-resistant acid phos-
phatase (TRACP) , sodium tartrate solution [3] should be added at 1/10 the volume
of the substrate solution. In both cases, the prepared reagents should be stored at
-20 ℃ , and used within 1 week.
VI. Outline of Procedure
< Cell sample >
< Blood sample >
Cell washing
Serum・Plasma
Cell lysis with Extraction solution
dilutio
Selection of Buffer (for ACP or for ALP)
Enzyme reaction at 37 ℃ for 15 - 60 min.
< kinetic assay for ALP only >
< Endpoint assay >
Stop reaction (color formation)
405 nm absorbance
3
TRACP & ALP Assay Kit
Cat. #MK301
v0804
URL:http://www.takara-bio.com