Acid phosphatase 100 μ l/ml preparation:
2
1
~ 2
15
stepwise dilution
μ l/ml
405 nm
50
4.014
25
3.993
12.5
3.995
6.26
4.066
3.125
4.011
1.5625
4.032
0.78125
3.411
0.390625
2.043
0.195313
1.147
0.097656
0.599
0.048828
0.356
0.024414
0.215
0.012207
0.157
0.006104
0.120
0.003052
0.108
0
0.084
(4) Measurement with Alkaline Phosphatase (ALP) as standard sample
A standard curve was made using 3 types of alkaline phosphatase, BAP (Cat.#
2120A, Lot K2601EA), CIAP (Cat. # 2250A, Lot E2301AB), or SAP (Cat.# 2660A, Lot
N301CB) of TAKARA BIO INC.
The 2-fold dilution series of the enzyme solutions, which be added Buffer for ALP
in the kit to each enzyme, was prepared with Extraction solution in the kit. Both
50 μ l of enzyme solution and 50 μ l of pNPP substrate dissolved in the Buffer for
ALP were added and mixed in one well of 96-well plate, and incubated at 37 ℃ for
30 minutes. Immediately after stopped by adding 50 μ l of Stop solution, absor-
bance at 405 nm of the 96-well plate was measured using a plate reader.
< Result >
All alkaline phosphatase (BAP, CIAP, and SAP) from TAKARA BIO were available as a
positive control for the kit.
(5) Comparison of Tartrate-resistant Acid Phosphate (TRACP) and Alkaline Phospha-
tase (ALP) by he preparation method of blood samples.
< Method >
Blood was collected from three rabbits simultaneously and the collocted blood
samples were preared by three methods (citrate plasma (PPP) , serum and hemoly-
sis serum) . 50 μ l of serial dilution samples and 50 μ l of the respective substrate
solution were mixed and reacted for 30 minutes at 37 ℃ . After adding 50 μ l of
stop solution, absorbance at 405 nm was measured using a plate reader. All of the
samples were measured simultaneously on the day when blood samples were
prepared.
Acid phosphatase standard
μ g/ml
8
TRACP & ALP Assay Kit
Cat. #MK301
v0804
URL:http://www.takara-bio.com