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LabAssay (TM) ALP

Manufacturer :
FUJIFILM Wako Shibayagi Corporation
Storage Condition :
Keep at 2-10 degrees C.
  • Structural Formula
  • Label
  • Packing
Comparison
Product Number
Package Size
Price
Inventory
Distributor
633-51021
Manufacturer
LABALP-M1
500Tests
List Price
JPY 21,000

In stock in Japan

Document

Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Overview

Introduction

Alkaline Phosphatase (ALP) is distributed in a variety of tissues such as liver, bone, and small intestine in animals. The change of the enzyme activity in tissues is an important hallmark for physiological phenomena as osteogenesis and so on. This kit is for Alkaline Phosphatase assay in a simultaneous multi-sample assay format with a microplate using p-Nitrophenylphosphate as a substrate.

Assay principle

p-Nitrophenylphosphate is hydrolyzed into p-Nitrophenol and Phosphoric Acid in the carbonate buffer (pH 9.8) in the presence of Alkaline Phosphatase in sample. Released p-Nitrophenol showing yellow color is optically measured at 405 nm wavelength as the enzyme activity.

04093371_img01.png
  • Performance

    Sample Serum (Mouse, Rat, Human)
    Standard curve range 0.0625 ~ 0.5 mmol/L
    Measurement time Approx. 20 min.
    Amount of sample 20 μL
    Measurement wavelength 405 nm
  • Standard Curve

    04093371_img02.png

Procedure

Perform the assay in the wells according to the following table scheme.

Test Standard Blank
Working Assay Solution 100 μL 100 μL 100 μL
Sample Sample 20 μL Dilution series 20 μL Distilled water 20 μL
Shake for 1 minute by plate mixer and incubate for 15 minutes at 37 ℃
Stop Solution 80 μL 80 μL 80 μL
Shake for 1 minute by plate mixer and measure the absorbance at 405 nm of wavelength using microplate reader.

Data

Spike-and-Recovery Test

Spike and Recovery Test of ALP, a known amount of standard is spiked into in human,mouse,rat serum sample n=2.

  • Human serum

    Spike (mmol/L) Mean (mmol/L) Recovery (mmol/L) Recovery(%)
    - 0.429 - -
    0.097 0.527 0.098 101.0
    0.197 0.587 0.158 80.2
    0.297 0.672 0.243 81.8
    0.507 0.866 0.437 86.2
  • Mouse serum

    Spike (mmol/L) Mean (mmol/L) Recovery (mmol/L) Recovery(%)
    - 0.284 - -
    0.060 0.343 0.059 98.3
    0.127 0.399 0.115 90.6
    0.251 0.507 0.223 88.8
    0.500 0.753 0.469 93.8
  • Rat Serum

    Spike (mmol/L) Mean (mmol/L) Recovery (mmol/L) Recovery(%)
    - 0.374 - -
    0.060 0.439 0.065 108.3
    0.127 0.483 0.109 85.8
    0.251 0.598 0.224 89.2
    0.500 0.855 0.481 96.2

Linearity-of-Dilution Test

  • Human serum
    04093371_img03.png
  • Mouse serum
    04093371_img04.png
  • Rat Serum
    04093371_img05.png

Example of measurement

Human serum, pooled, male
No. Measured(mmol/L) Mean(mmol/L) SD CV(%)
1 0.558 0.555 0.556 0.003 0.5
Human plasma K2EDTA, pooled, male
No. Measured(mmol/L) Mean(mmol/L) SD CV(%)
1 0.070 0.071 0.070 0.001 1.2
  • 20% Diluted Normal Mouse serum, CD-1(ICR), male
    No. Measured(mmol/L) Mean(mmol/L) SD CV(%)
    1 0.320 0.301 0.310 0.013 4.2
    2 0.452 0.454 0.453 0.001 0.2
    3 0.505 0.484 0.495 0.015 3.0
    4 0.433 0.451 0.442 0.013 2.9
    5 0.372 0.375 0.373 0.002 0.5
  • Normal Mouse plasma K2EDTA, CD-1(ICR), male
    No. Measured(mmol/L) Mean(mmol/L) SD CV(%)
    1 0.081 0.079 0.080 0.001 1.5
    2 0.213 0.202 0.208 0.008 3.8
    3 0.099 0.098 0.098 0.000 0.1
    4 0.056 0.061 0.059 0.004 6.5
    5 0.110 0.109 0.109 0.001 0.6
  • 10% Diluted Normal Rat serum, Sprague Dawley, male
    No. Measured(mmol/L) Mean(mmol/L) SD CV(%)
    1 0.541 0.534 0.538 0.005 1.0
    2 0.434 0.448 0.441 0.010 2.2
    3 0.357 0.357 0.357 0.000 0.0
    4 0.461 0.458 0.460 0.002 0.5
    5 0.308 0.293 0.301 0.011 3.6
  • Normal Rat plasma K2EDTA, Sprague Dawley, male
    No. Measured(mmol/L) Mean(mmol/L) SD CV(%)
    1 0.170 0.172 0.171 0.001 0.7
    2 0.145 0.134 0.139 0.007 5.3
    3 0.262 0.257 0.260 0.004 1.4
    4 0.209 0.202 0.205 0.006 2.7
    5 0.174 0.171 0.172 0.002 1.2
  • Normal Canine serum, Beagle, male
    No. Measured(mmol/L) Mean (mmol/L) SD CV(%) Age(years)
    1 0.395 0.385 0.390 0.005 1.2 4
    2 0.138 0.138 0.138 0.000 0.2 6
    3 0.314 0.303 0.309 0.005 1.7 6
    4 0.323 0.299 0.311 0.012 3.8 11
    5 0.323 0.326 0.325 0.002 0.5 12
    6 0.297 0.290 0.293 0.004 1.2 6
    7 0.535 0.535 0.535 0.000 0.0 10
    8 0.297 0.295 0.296 0.001 0.5 4
  • Normal Canine plasma EDTA, Beagle, male
    No. Measured(mmol/L) Mean (mmol/L) SD CV(%) Age(years)
    1 0.025 0.031 0.028* 0.003 10.7 4
    2 0.030 0.033 0.032* 0.002 4.8 6
    3 0.030 0.036 0.033* 0.003 8.6 10
    4 0.056 0.069 0.062* 0.006 10.2 12
    5 0.058 0.061 0.059* 0.002 2.8 6
    6 0.026 0.028 0.027* 0.001 3.8 11
    7 0.021 0.022 0.022* 0.001 3.1 10
    8 0.065 0.070 0.067 0.003 3.7 12
  • Normal Feline serum, Short hair, male
    No. Measured(mmol/L) Mean (mmol/L) SD CV(%) Age(years)
    1 0.204 0.196 0.200 0.004 2.1 3
    2 0.158 0.165 0.162 0.004 2.2 3
    3 0.221 0.213 0.217 0.004 1.8 6
    4 0.229 0.234 0.232 0.003 1.2 4
    5 0.297 0.295 0.296 0.001 0.3 6
    6 0.155 0.149 0.152 0.003 2.0 6
    7 0.398 0.388 0.393 0.005 1.2 1
    8 0.343 0.333 0.338 0.005 1.4 1
  • Normal Feline plasma EDTA, Short hair, male
    No. Measured(mmol/L) Mean (mmol/L) SD CV(%) Age(years)
    1 0.227 0.227 0.227 0.000 0.0 0.5
    2 0.050 0.047 0.048* 0.002 3.4 1
    3 0.086 0.088 0.087 0.001 1.0 3
    4 0.107 0.103 0.105 0.002 2.1 0.5
    5 0.138 0.136 0.137 0.001 0.7 3
    6 0.062 0.058 0.060* 0.002 3.3 3
    7 0.031 0.028 0.030* 0.001 4.0 0.5
    8 0.057 0.056 0.056* 0.000 0.6 1

References

  1. Ito, S. et al. : J. Pharmacol. Exp. Ther., 333, 341(2010). *Extraction liqiud of mouse kidney
  2. Matsuyama, A. et al. : Clin. Exp. Pharmacol. Physiol., 45, 75(2018). *MC3T3-E1cell, C2C12Ccell
  3. Chiba, T. et al. : J. Atheroscler. Thromb, 23, 1099(2016). *Mouse plasma
  4. Kohno, Y. et al. : Stem Cell Res. Ther., 8, 115(2017). *Mesenchymal stem cell

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