![background image](http://html.mh-extra.com/html/takara/mk301/mk301_manual_814465009.webp)
< Results >
< TRACP activity >
A
405
nm absorbanse
TRACP citrate plasma( PPP )
serum
hemolysis serum
ID No.
x 20
x 40
x 80
x 20
x 40
x 80
x 20
x 40
x 80
Rb No. 1 0.821 0.420 0.247 0.834 0.487 0.309 0.834 0.487 0.300
Rb No. 2 1.045 0.520 0.311 0.704 0.422 0.268 1.066 0.582 0.360
Rb No. 3 0.702 0.370 0.237 1.000 0.579 0.353 0.768 0.360 0.275
< ALP activity >
A
405
nm absorbanse
ALP
citrate plasma( PPP )
serum
hemolysis serum
ID No.
x 2
x 4
x 8
x 2
x 4
x 8
x 2
x 4
x 8
Rb No. 1 0.514 0.321 0.217 0.801 0.469 0.295 1.074 0.599 0.374
Rb No. 2 0.481 0.299 0.206 0.528 0.327 0.217 0.801 0.483 0.304
Rb No. 3 0.348 0.231 0.165 0.718 0.431 0.275 0.650 0.398 0.252
As the phosphate activity is different depending on the preparation method of
blood sample, it is necessary to use the sample of the same preparation method.
(6) Influence of freeze-thaw cycles of blood samples on the phosphate activity.
< Method >
Serum samples were collected from three rabbits. Samples were devided into four
portion and each sample was repeated various cycles of freeze-thawing ( - 80 ℃
⇔ 25 ℃)
For the measurement of TRACP (tartrate-resistant acid phosphate), samples were
diluted by 20-, 40- and 80- fold. For the measurement of ALP (alkaline phosphate),
samples were diluted by 2-, 4- and 8- fold.
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 solu-
tion, absorbance at 405 nm was measured using a plate reader. All of the samples
were measured simultaneously on the day when serum samples were prepared.
< Results >
< TRACP activity >
A
405
nm absorbanse
TRACP
no freezing
freezing-thaw 1 cycle freezing-thaw 2 cycles freezing-thaw 3 cycles
ID No.
x 20 x 40 x 80 x 20 x 40 x 80 x 20 x 40 x 80 x 20 x 40 x 80
Rb No. 1 0.826 0.477 0.295 0.832 0.438 0.284 0.778 0.452 0.292 0.736 0.432 0.277
Rb No. 2 1.087 0.577 0.342 1.078 0.573 0.343 1.021 0.580 0.353 0.980 0.565 0.344
Rb No. 3 0.743 0.409 0.252 0.754 0.429 0.269 0.749 0.434 0.279 0.710 0.424 0.275
< ALP activity >
A
405
nm absorbanse
ALP
no freezing
freezing-thaw 1 cycle freezing-thaw 2 cycles freezing-thaw 3 cycles
ID No.
x 2
x 4
x 8
x 2
x 4
x 8
x 2
x 4
x 8
x 2
x 4
x 8
Rb No. 1 0.616 0.338 0.281 0.602 0.328 0.212 0.592 0.335 0.214 0.588 0.330 0.221
Rb No. 2 0.577 0.327 0.206 0.600 0.338 0.214 0.562 0.324 0.209 0.577 0.334 0.213
Rb No. 3 0.400 0.243 0.164 0.423 0.255 0.170 0.440 0.261 0.175 0.415 0.248 0.173
It seemed that both of TRACP and ALP activities were not comparatively influ-
enced by freeze-thaw cycles of blood samples, but it was preferable even twice of
freeze-thaw cycles in the same condition.
9
TRACP & ALP Assay Kit
Cat. #MK301
v0804
URL:http://www.takara-bio.com