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  2. Life Science
  3. Metabolism Research
  4. ELISA / Assay Kits (Metabolism)
  5. Creatinine Assay Kits
  6. LabAssay (TM) Creatinine

LabAssay (TM) Creatinine

for Cellbiology
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
GHS :
Citations (1)
PubMed Central® search
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SDS
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Product Number
Package Size
Price
Inventory
Distributor
291-93901
Barcode No
4548995102515
500Tests
List Price
JPY 33,000

In stock in Japan

Please check here for notes on products and prices.

Document

SDS
Product Specification Sheet
Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate
Analytical Charts

Kit component

For 500 tests

Deproteinizing Reagent 150 mL/1 bottle
Picric Acid Reagent 50 mL/1 bottle
0.75 mol/L Sodium Hydroxide Solution 50 mL/1 bottle
Standard Solution 15 mL/1 bottle

Product Overview

Creatinine is a metabolic product formed by the dehydration and cyclization of creatine and is a type of non-protein nitrogen. The produced creatinine is excreted in the urine via the kidneys. The serum creatinine level rises when kidney function is compromised.

LabAssay™ Creatinine is a kit designed to measure creatinine in samples using the Jaffé reaction. With the use of a microplate, this kit provides a convenient method for measuring creatinine in samples.

[Note] LabAssay™ series are reagents for research purposes. They cannot be used for diagnostic purposes.

Kit Performance

Analysis sample Human Serum/Plasma
Mouse Serum/Plasma/Urine
Rat Serum/Plasma
Dog Serum/Plasma
Cat Serum/Plasma
Calibration curve range 2.5-10 mg/dL
Sample volume 50 μL
Measurement duration Approx. 40 min
Wavelength 520 nm

Example of Calibration Curve

Example of Calibration Curve

Assay Principle

A deproteinizing reagent is added to the sample and centrifuged, and the supernatant is obtained. When picric acid is added to the supernatant, the creatinine in the sample reacts with the picric acid in an alkaline solution to produce an orange-red condensation product (Jaffé reaction). By measuring the absorbance of this orange-red color, the creatinine concentration in the sample is determined.

Procedure

Assay in a microplate
Perform the assay in the wells according to the following table scheme.

Test Standard Blank
First operations should be performed in the sample tubes.
Sample Sample
50 μL		 Standard solution
50 μL		 Water
50 μL
Deproteinizing reagent 300 μL 300 μL 300 μL
Mix well, and stand at room temperature for 10 min.
Centrifuge over 2500 rpm for 10 min.
Subsequent operations should be performed in the wells.
Supernatant 100 μL 100 μL 100 μL
　 Stand at 25 ℃ - 30 ℃
Picric Acid Reagent 50 μL 50 μL 50 μL
0.75 mol/L Sodium Hydroxide Solution 50 μL 50 μL 50 μL
　 Stand at 25 ℃ - 30 ℃ for 20 min.
Measure the absorbance of the test sample and standard solution
at 520 nm with the blank solution as the control within the next 30 min.

Data

Repeatability (within run precision)

Repeatability was conducted by measuring sixfold assays of serum samples from human, mouse and rat at two concentrations with this product.

n＼ID Human serum Mouse serum Rat serum
ID1 (mg/dL) ID2 (mg/dL) ID1 (mg/dL) ID2 (mg/dL) ID1 (mg/dL) ID2 (mg/dL)
1 5.67 2.99 4.27 2.86 4.85 2.6
2 5.40 2.91 4.76 2.67 4.63 2.37
3 5.82 3.34 4.59 3.01 4.95 2.69
4 5.47 3.07 4.18 2.64 4.55 2.37
5 5.26 2.95 4.59 2.53 4.48 2.25
6 5.78 3.35 4.84 2.57 4.99 2.54
mean 5.57 3.10 4.54 2.71 4.74 2.47
SD 0.224 0.196 0.263 0.185 0.217 0.166
CV(%) 4.0 6.3 5.8 6.8 4.6 6.7

[Result]
The CV (%) of human serum was 4.0-6.3%, mouse serum was 5.8-6.8% and rat serum was 4.6-6.7%, indicating good repeatability.

Reproducibility (between run precision)

Reproducibility was conducted by measuring serum samples from human, mouse and rat at three concentrations each for four days with this product.

Day＼ID Human serum Mouse serum Rat serum
ID3 (mg/dL) ID4 (mg/dL) ID5 (mg/dL) ID3 (mg/dL) ID4 (mg/dL) ID5 (mg/dL) ID3 (mg/dL) ID4 (mg/dL) ID5 (mg/dL)
1 5.78 5.13 2.76 6.30 5.72 3.40 6.60 5.52 2.69
2 6.07 5.42 2.84 6.42 6.09 3.57 6.49 5.87 2.86
3 6.23 5.60 2.98 6.55 5.89 3.32 6.96 6.31 2.91
4 6.04 5.30 2.67 6.27 5.86 3.52 6.76 6.10 2.84
mean 6.03 5.36 2.81 6.39 5.89 3.45 6.70 5.95 2.83
SD 0.186 0.198 0.131 0.128 0.153 0.114 0.204 0.338 0.0947
CV(%) 3.1 3.7 4.7 2.0 2.6 3.3 3.0 5.7 3.4

[Result]
The CV (%) of human serum was 3.1-4.7%, mouse serum was 2.0-3.3% and rat serum was 3.0-5.7%, indicating good reproducibility.

Dilution Linearity Test

Human, mouse, and rat samples, were serially diluted twofold with saline and measured in duplicate to assess linearity.

Human sample
Mouse sample
Rat sample

[Result]
All measurement results showed good linearity within the measurement range.

Spike and Recovery Test

Standard solutions at three concentrations were added to serum/plasma (EDTA, heparin) from human, mouse, rat. Next, spike recovery tests were conducted, with each measurement performed in duplicate.

Human sample

Spiked amount
(mg/dL)		 Measurement value
(mg/dL)		 Recovery volume
(mg/dL)		 Recovery rate
(%)
Serum - 3.68 - -
2.71 6.37 2.70 99.6
3.55 7.10 3.42 96.3
4.21 7.55 3.87 91.9
Average 96.0
Plasma
(EDTA)		 - 3.43 - -
3.30 6.47 3.04 92.1
4.33 7.39 3.96 91.5
5.13 7.69 4.26 83.0
Average 88.9
Plasma
(heparin)		 - 3.37 -
3.31 6.97 3.60 109
4.34 7.86 4.49 103
5.14 8.21 4.84 94.2
Average 102

Mouse sample

Spiked amount
(mg/dL)		 Measurement value
(mg/dL)		 Recovery volume
(mg/dL)		 Recovery rate
(%)
Serum - 2.68 - -
3.00 5.90 3.22 107
5.14 8.08 5.40 105
6.74 8.90 6.22 92.3
Average 102
Plasma
(EDTA)		 - 3.14 - -
2.71 5.63 2.49 91.9
4.65 7.74 4.60 98.9
6.10 8.25 5.11 83.8
Average 91.5
Plasma
(heparin)		 - 3.01 -
2.85 5.48 2.47 86.7
4.88 7.74 4.73 96.9
6.41 8.60 5.59 87.2
Average 90.3

Rat sample

Spiked amount
(mg/dL)		 Measurement value
(mg/dL)		 Recovery volume
(mg/dL)		 Recovery rate
(%)
Serum - 2.86 - -
3.27 5.61 2.75 84.1
5.60 8.13 5.27 94.1
7.35 9.34 6.48 88.2
Average 88.8
Plasma
(EDTA)		 - 2.78 - -
2.79 5.22 2.44 87.5
4.78 8.30 5.52 115
6.27 9.22 6.44 103
Average 102
Plasma
(heparin)		 - 2.88 -
3.17 5.53 2.65 83.6
5.43 8.25 5.37 98.9
7.13 9.27 6.39 89.6
Average 90.7

[Result]
Good recovery rates were confirmed.

Measurement Using Actual Samples

Human sample

ID＼n Measurement value
(mg/dL)		 mean SD CV
(%)
1 2
Serum ID1 3.69 3.67 3.68 0.0120 0.3
Plasma (EDTA) ID1 3.43 3.44 3.43 0.00707 0.2

Rat sample (S.D. rat)

ID＼n Measurement value
(mg/dL)		 mean SD CV
(%)
1 2
Serum ID1 0.553 0.546 0.550 0.00495 0.9
ID2 0.716 0.824 0.770 0.0764 9.9
ID3 1.12 1.26 1.19 0.0948 8.0
ID4 1.22 1.29 1.25 0.0502 4.0
ID5 1.035 0.994 1.01 0.0290 2.9
Plasma
(EDTA)		 ID1 0.319 0.288 0.304 0.0219 7.2
ID2 0.638 0.627 0.633 0.00778 1.2
ID3 0.824 0.871 0.848 0.0332 3.9
ID4 0.661 0.681 0.671 0.0141 2.1
ID5 0.205 0.233 0.219 0.0198 9.0

Mouse sample (CD-1(ICR))

ID＼n Measurement value
(mg/dL)		 mean SD CV
(%)
1 2
Serum ID1 0.199 0.205 0.202 0.00424 2.1
ID2 0.492 0.507 0.500 0.0106 2.1
ID3 0.734 0.722 0.728 0.00849 1.2
ID4 0.641 0.716 0.679 0.0530 7.8
ID5 0.192 0.219 0.206 0.0191 9.3
Plasma
(EDTA)		 ID1 0.095 0.103 0.0990 0.00566 5.7
ID2 0.607 0.559 0.583 0.0339 5.8
ID3 0.556 0.528 0.542 0.0198 3.7
ID4 0.490 0.546 0.518 0.0396 7.6
ID5 0.113 0.110 0.112 0.00212 1.9

Dog sample (Beagle)

ID＼n Measurement value
(mg/dL)		 mean SD CV
(%)
1 2
Serum ID1 1.67 1.48 1.58 0.134 8.5
ID2 1.16 1.07 1.12 0.0636 5.7
ID3 1.54 1.41 1.48 0.0919 6.2
ID4 1.58 1.40 1.49 0.127 8.5
ID5 1.46 1.49 1.48 0.0212 1.4
ID6 1.17 1.20 1.19 0.0212 1.8
ID7 1.16 1.20 1.18 0.0283 2.4
ID8 1.23 1.30 1.27 0.0495 3.9
Plasma
（EDTA）		 ID1 1.48 1.45 1.47 0.0212 1.4
ID2 1.21 1.18 1.20 0.0212 1.8
ID3 1.33 1.22 1.28 0.0778 6.1
ID4 1.56 1.49 1.53 0.0495 3.2
ID5 1.31 1.29 1.30 0.0141 1.1
ID6 1.04 1.04 1.04 0.00 0.0
ID7 1.14 1.16 1.15 0.0141 1.2
ID8 1.26 1.29 1.28 0.0212 1.7

Cat sample (Short hair)

ID＼n Measurement value
(mg/dL)		 mean SD CV
(%)
1 2
Serum ID1 1.86 1.78 1.82 0.0566 3.1
ID2 1.58 1.53 1.56 0.0354 2.3
ID3 1.81 1.68 1.75 0.0919 5.3
ID4 2.17 1.95 2.06 0.156 7.6
ID5 1.88 1.92 1.90 0.0283 1.5
ID6 1.89 1.86 1.88 0.0212 1.1
ID7 1.71 1.76 1.74 0.0354 2.0
ID8 1.48 1.56 1.52 0.0566 3.7
Plasma
（EDTA）		 ID1 1.63 1.61 1.62 0.0141 0.9
ID2 1.41 1.36 1.39 0.0354 2.6
ID3 1.78 1.62 1.70 0.113 6.7
ID4 1.97 1.76 1.87 0.148 8.0
ID5 1.66 1.67 1.67 0.00707 0.4
ID6 1.28 1.29 1.29 0.00707 0.6
ID7 1.26 1.30 1.28 0.0283 2.2
ID8 1.60 1.71 1.66 0.0778 4.7

FAQ

About sample

Do anticoagulants, glycolysis inhibitors, or urinary preservatives affect the assay?
Anticoagulants such as heparin, citrate, oxalate, and EDTA, as well as the glycolysis inhibitor sodium fluoride, do not affect the assay when used at standard concentrations. Toluene is recommended as a preservative for urine samples.
Is there any influence from substances that interfere with measurement (ascorbic acid, bilirubin, etc.)?
Hemolysis, ascorbic acid and bilirubin have almost no effect on the measured values. However a substance with an active methylene group causes an increase of the value.

About kit usage

What instruments, and equipment are required for the assay using this kit?
The instruments and equipment required for the use of this kit are listed below.
  • 96 well microplate (transparent type)
  • Micropipette
  • Plate mixer*
  • Incubator maintaining at 25-30°C*
  • Microplate reader with 520 nm wavelength filter

(*This is not necessary depending on the model of the microplate reader.)

(For test tube method)

  • Test tube
  • Pipette
  • Spectrophotometer or colorimeter with 500-530 nm wavelength filter

Overview / Applications

Outline Creatinine is a metabolite which is produced by creatine phosphate directly or dehydration of creatine in muscle and nerve. It is excreted from the body through kidney glomerular filtration. This product can be used to measure the creatinin levels in samples by Jaffe's reaction where creatinine produces quantitatively an orange color with picric acid in alkaline condition.

Property

Manufacturer Information

Alias

  • LabAssay
    636-51011 (Previous code)
    LABCREA-M1 (Previous code)
    290-65901 (Previous code)

For research use or further manufacturing use only. Not for use in diagnostic procedures.

Product content may differ from the actual image due to minor specification changes etc.

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