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Caffeine ELISA Kit Wako

for Immunochemistry
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
GHS :
  • Structural Formula
  • Label
  • Packing
SDS
Comparison
Product Number
Package Size
Price
Inventory
Distributor
296-85901
Barcode No
4548995097286
96Tests
List Price
JPY 85,000

In stock in Japan

Document

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

Kit component

96 tests

Antibody-coated Plate 1 plate
Caffeine Standard 100 μL
Buffer 60 mL
Peroxidase-conjugated Caffeine Solution 50 μL
TMB Solution 12 mL
Stop Solution 12 mL
Wash Solution (10x) 100 mL
Plate Seal 4 seals

Product Overview

Caffeine is a type of alkaloid found in coffee beans and tea leaves. It induces wakefulness and has an excitatory effect through the competitive inhibition of adenosine receptors. Caffeine is also known to have various physiological effects, including vasoconstrictive, cardiotonic, and diuretic effects, as well as promoting gastric secretion. Reports have shown that blood caffeine can be a useful biomarker for Parkinson's disease,1) and that caffeine has protective effects against Parkinson's disease.2) Therefore, it is attracting attention in the field of neurological and psychiatric disorders.

Caffeine is mainly measured by LC-MS/MS or HPLC, but the assay of caffeine used to include the need for expensive analytical equipment and complicated pretreatment. Fujifilm Wako has developed an ELISA kit that can measure the concentration of caffeine in human saliva, serum or plasma and mouse/rat serum or plasma.

Features

  • Not need for expensive analytical equipment like LC-MS/MS and HPLC
  • Easy-to-use pretreatment
  • Can be used to measure caffeine in human saliva, serum or plasma and mouse/rat serum or plasma.

Assay Principle

04520963_img01.png

This kit is based on a competitive assay, and absorbance intensity decreases as the amount of caffeine in the sample increases.

Kit Performance

Calibration curve range 0.244 - 1,000 ng/mL
Assay target Caffeine
Analysis sample Human saliva/serum/plasma (EDTA/heparin)
Mouse serum/plasma
Rat serum/plasma
Sample volume 55 μL (2-fold dilution, duplicate)
Measurement duration Approx. 2 hours 20 minutes
Detection method Chromogenic
(Primary wavelength 450 nm / reference wavelength 620 nm)

Example of Calibration Curve

04520963_img02.png

Data

Spiked recovery test

Human

Amount
spiked
(ng/mL)
Measured
value
(ng/mL)
Recovery
volume
(ng/mL)
Recovery
rate
(%)
Saliva 1 0 11.3 - -
10 21.1 9.80 98.0
50 64.7 53.4 107
250 268 257 103
Average 103
Saliva 2 0 3.15 - -
10 13.8 10.7 107
50 53.0 49.9 99.8
250 276 273 109
Average 105
Saliva 3 0 7.79 - -
10 18.2 10.4 104
50 58.3 50.5 101
250 267 259 104
Average 103
Serum 0.00 1.31
1.00 2.33 1.02 102
50.0 49.9 48.6 97.2
300 295 294 98.0
Average 99.1
Plasma
(EDTA)
0.00 2.11
1.00 3.15 1.04 104
50.0 45.5 43.4 86.8
300 289 287 95.7
Average 95.5
Plasma
(Heparin)
0.00 3.21
1.00 4.32 1.11 111
50.0 54.9 51.7 103
300 312 309 103
Average 106

Mouse

Amount
spiked
(ng/mL)
Measured
value
(ng/mL)
Recovery
volume
(ng/mL)
Recovery
rate
(%)
Serum 0.00 9.72 - -
1.00 10.7 0.980 98.0
50.0 62.6 52.9 106
300 314 304 101
Average 102
Plasma 0.00 8.88 - -
1.00 9.87 0.990 99.0
50.0 58.3 49.4 98.8
300 317 308 103
Average 100

Rat

Amount
spiked
(ng/mL)
Measured
value
(ng/mL)
Recovery
volume
(ng/mL)
Recovery
rate
(%)
Serum 0.00 17.1 - -
1.00 18.1 1.00 100
50.0 66.1 49.0 98.0
300 322 305 102
Average 100
Plasma 0.00 4.23 - -
1.00 5.22 0.990 99.0
50.0 49.9 45.7 91.4
300 322 318 106
Average 98.8

Dilution linearity test

Human

04520963_img06.png

 

04520963_img03.png

Mouse

04520963_img04.png

Rat

04520963_img05.png

References

FAQ

About sample

How should samples be stored?
For long-term storage of samples, freezing at -35°C or below is recommended. Avoid repeated freezing and thawing. Frozen samples should be thawed just before the assay and thoroughly mixed. For saliva samples, assay results did not change significantly after storage at room temperature for 6 hours, in a refrigerator for 24 hours at -20°C and -80°C for 1 month. (Fluctuations in room temperature were within ±10% of the temperature at hour 0.) Fujifilm Wako has also observed that assay results did not change significantly within five cycles of freezing and thawing.
What is the recommended method of collection of saliva samples?
The passive drool method is recommended for the collection of saliva samples. (the passive drool method requires subjects to salivate into a vial or tube.) It has been confirmed that samples collected using cotton swabs or inert polymer swabs can be used similarly to samples collected by the passive drool method. However, caution should be taken because some swab materials may absorb target analytes.

About the kit

What reagents, instruments, and equipment are required for the assay using this kit?
  • Purified water (distilled water)
  • Tubes for dilution of standard solutions and samples
  • Glassware for dilution of Wash Solution (graduated cylinders, beakers)
  • Micropipettes (for 10μL disposable tips and 200 to 500 μL disposable tips)
  • Continuous dispensing pipette, capable of dispensing 100 μL continuously (preferred if available)
  • Paper towels or other absorbent material (to remove liquid left on the plate after washing)
  • Vortex-type mixer
  • Microplate shaker (range approx. 600 to 800 rpm)
  • Automatic washer for 96-well plate (if available) or washing bottle
  • Microplate reader for 96-well plates (for absorbance measurement)
  • Software for data processing
Is it possible to purchase the antibody provided in this kit?
This antibody are not sold separately.
Can I divide plates?
Since the plate in this kit are separable and each row (8 wells) is detachable, divided use of the kit is possible.

Overview / Applications

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Manufacturer Information

Alias

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

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