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LabAssay ™ NEFA

Manufacturer :
FUJIFILM Wako Shibayagi Corporation
Storage Condition :
Keep at 2-10 degrees C.
Molecular Weight :
0
GHS :
  • Structural Formula
  • Label
  • Packing
Comparison
Product Number
Package Size
Price
Inventory
Distributor
633-52001
Manufacturer
LABNEFA-M1
500Tests
List Price
408.00 USD

Out of Stock

Document

Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Kit component

Kit component

Chromogen Reagent A for 10 mL x 4 vials
Solvent A 45 mL x 1 vial
Chromogen Reagent B for 20 mL x 4 vials
Solvent B 90 mL x 1 vial
Standard Solution  (1mEq/L of Oleic acid) *amount of oleic acid; 1mEq=1mmol 7 mL x 1 vial

Overview

※For research purposes only with animal serum/plasma, Not for Diagnostic Use.

Introduction

NEFA (Non‐esterified fatty acid) in the blood is transported complexed with an albumin to peripheral tissues. They are important sources of fuel for the peripheral tissues. The concentration of NEFA in the blood is regulated by a release from the adipose tissues, a consumption in the periphral tissues or a take up from the liver.

Assay principle

NEFA (Non‐esterified fatty acid) forms Acyl-CoA in the presence of Acyl-CoA synthetase (ACS). Hydrogen peroxide, which is produced by a reaction between the Acyl-CoA and Acyl-CoA oxidase (ACOD), promotes oxidative condensation of 3-methyl-N-ethyl-N-(β-hydroxyethyl)-aniline (MEHA) with 4-aminoantipyrine. LabAssay™ NEFA can be used for the quantitative determination of NEFA in samples by measuring absorbance of a blue purple color which is generated by the oxidative condensation reaction.

  • Performance

    Sample Serum, plasma(Mouse, Rat, dog, cat)
    Standard curve range 0.4~1.97 mEq/L
    Measurement time approx. 20 min.
    Amount of sample 4 μL
    Measurement wavelength 550 nm
  • Standard Curve

    04200685_img01.png

Procedure

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

No. Standard Solution Distilled or deionized water Sample volume Concentration
1 80 µL 120 µL 4 μL 0.40 mEq/L
2 120 µL 80 µL 4 μL 0.60 mEq/L
3 160 µL 40 µL 4 μL 0.80 mEq/L
4 Undiluted - 4 μL 1.00 mEq/L
5 Undiluted - 8 μL 1.97 mEq/L*1

*1: The test sample volume is usually 4 μL, but 8 μL is taken in this case. The NEFA concentration must be corrected accordingly as indicated in the table above.

Data

Spike-and-Recovery Test

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

  • Rat Sample

    Spike
    (mEq/L)
    Mean
    (mEq/L)
    Recovery
    (mEq/L)
    Recovery
    (%)
    Serum - 0.57    
    0.40 1.00 0.43 106.4%
    0.61 1.21 0.65 106.2%
    0.78 1.40 0.83 106.4%
    1.01 1.57 1.01 99.7%
    Plasma
    (EDTA)
    - 0.41    
    0.40 0.86 0.45 111.5%
    0.61 1.07 0.66 108.5%
    0.78 1.27 0.86 109.5%
    1.01 1.44 1.02 101.4%
  • Mouse Sample

    Spike
    (mEq/L)
    Mean
    (mEq/L)
    Recovery
    (mEq/L)
    Recovery
    (%)
    Serum - 0.70    
    0.40 1.07 0.37 91.7%
    0.61 1.31 0.61 100.1%
    0.78 1.44 0.74 94.5%
    1.01 1.62 0.92 91.3%
    Plasma
    (EDTA)
    - 0.62    
    0.40 0.99 0.37 92.5%
    0.61 1.29 0.68 111.5%
    0.78 1.38 0.77 97.8%
    1.01 1.62 1.00 99.1%

Linearity-of-Dilution Test

  • Rat sample

    04200685_img03.png
  • Mouse sample

    04200685_img04.png
  • D-MEM/10% FBS

    04200685_img05.png

Examples of measurement

  • 20% Diluted Nomal Mouse serum, CD-1(ICR), male
    No. Mean (mEq/L) SD CV(%)
    1 1.118 0.001 0.1
    2 0.788 0.007 0.9
    3 0.715 0.022 3.1
    4 0.794 0.002 0.2
    5 0.829 0.005 0.6
  • Nomal Mouse plasma K2EDTA, CD-1(ICR), male
    No. Mean SD CV(%)
    1 *0.309 0.002 0.6
    2 *0.320 0.002 0.6
    3 *0.205 0.004 2.0
    4 *0.179 0.008 4.6
    5 *0.305 0.017 5.6
  • 10% Diluted Nomal Rat serum, Sprague Dawley, male
    No. Mean SD CV(%)
    1 *0.300 0 0.0
    2 *0.371 0.004 1.1
    3 *0.289 0.017 5.8
    4 *0.365 0 0.1
    5 *0.382 0.029 7.6
  • Nomal Rat plasma K2EDTA, Sprague Dawley, male
    No. Mean SD CV(%)
    1 *0.333 0.004 1.2
    2 *0.356 0.003 0.8
    3 *0.325 0.020 6.0
    4 *0.218 0.005 2.6
    5 0.515 0.010 1.9
  • Nomal Canine serum, Beagle, male 
    No. Mean (mEq/L) SD CV(%) Age(years)
    1 0.656 0.0027 0.4 4
    2 0.665 0.0320 4.8 6
    3 0.425 0.0090 2.0 6
    4 *0.38 0.0057 1.5 11
    5 0.882 0.0070 0.8 12
    6 0.599 0.0060 1.1 6
    7 1.006 0.0267 2.7 10
    8 0.506 0.0060 1.2 4
  • Nomal Canine plasma EDTA, Beagle, male
    No. Mean (mEq/L) SD CV(%) Age(years)
    1 0.442 0.0033 0.7 4
    2 0.427 0.0110 2.6 6
    3 *0.308 0.0037 1.2 10
    4 0.478 0.0123 2.6 12
    5 0.473 0.0050 1.0 6
    6 *0.367 0.0047 1.2 11
    7 0.780 0.0133 1.7 10
    8 0.411 0.0387 9.4 12
  • Nomal Feline serum, Short hair, male
    No. Mean (mEq/L) SD CV(%) Age(years)
    1 0.717 0.0117 1.6 3
    2 0.635 0.0100 1.6 3
    3 1.004 0.0053 0.5 6
    4 0.981 0.0093 0.9 4
    5 0.636 0.0170 2.7 6
    6 1.193 0.0267 2.3 6
    7 1.022 0.0893 8.8 1
    8 *0.382 0.0157 4.1 1
  • Nomal Feline plasma EDTA, Short hair, male 
    No. Mean (mEq/L) SD CV(%) Age(years)
    1 1.099 0.0103 0.9 0.5
    2 *0.174 0.0033 2.0 1
    3 *0.241 0.0043 1.7 3
    4 *0.320 0.0067 2.2 0.5
    5 *0.359 0.0060 1.7 3
    6 *0.197 0.0120 6.2 3
    7 *0.215 0.0240 11.1 0.5
    8 *0.131 0.0160 12.3 1

References

Kobayashi, Y. et al. : J. Pharmacogn. Nat. Prod., online (2015). Effects of Morus alba L. (Mulberry) Leaf Extract in Hypercholesterolemic Mice on Suppression of Cholesterol Synthesis *Extraction liquid of mouse liver

Gao, F. et al. : Evid. Based Complement. Alternat. Med., 2015, 801291 (2015). Effect of Keishibukuryogan on Genetic and Dietary Obesity Models *Extraction liquid of rat liver

Ogawa, K. et al. : Reprod. Med. Biol., online (2018). Non-esterified fatty acid-associated ability of follicular fluid to support porcine oocyte maturation and development *Pig cumulus-oocyte complexes

Wang, F. et al. : J. Mol. Endocrinol., 52, 133 (2014). LGR4 acts as a link between the peripheral circadian clock and lipid metabolism in liver *Mouse plasma

Uchida, K. et al. : Exp. Anim., 58, 181 (2009). Hyperlipidemia and hyperinsulinemia in the spontaneous osteoarthritis mouse model, STR/Ort *Mouse serum

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