V.2/Oct/2018 4.
Materials required but not provided
Precautions
All procedures should be performed under contamination-free conditions to obtain reliable results.
Make sure to avoid cross-contaminations via equipment, devices, tubes, containers, pipette tips, etc.
Rapid Test ProⅡ for Egg (Cat.# M2261)
The use of disposable materials is recommended.
Rapid Test ProⅡ for Casein (Cat.# M2262)
Homogenizer / blender
Rapid Test ProⅡ for Gluten (Cat.# M2263) Scale capable of weighing 1g
Vortex mixer
Rapid Test ProⅡ for Buckwheat (Cat.# M2264)
Water bath*
Rapid Test ProⅡ for Peanut (Cat.# M2265) Centrifuge (for 3000 x g)*
Filter paper*
Rapid Test ProⅡ for Soya (Cat.# M2266) pH test strip
Heat-resistant glove
＊
The items may not be required depending on the test method or sample condition.
The Quick Detection for Protein of Allergenic Ingredients
in Foods and Food-processing Equipment
5. Sample Extraction
10 tests Precautions
Prior to use, bring all reagents to 20−30˚C (68−86˚F) and gently vortex the contents into a
For Research or Laboratory Use Only homogeneous solution.
Not for Use in Diagnostic Procedures Confirm and adjust the pH of Sample Extract close to neutral (pH 6−8) as required.
Please read full descriptions in this manual before use Wear suitable protective clothing, goggles and gloves when handling the kit.
Manufactured by:
A-1. For food sample【Recommended Extraction Method】
Morinaga Institute of Biological Science, Inc. (MIoBS)
1. Grind and mix the test food sample to homogeneity with a contamination-free homogenizer/
2-1-16 Sachiura, Kanazawa-ku, Yokohama 236-0003, Japan
blender.
E-mail: info@miobs.com
Website: http://www.miobs-e.com
2. Put 1.0 g of the homogenized sample in a polypropylene tube (L) and add 1 pack of Extraction
Solution.
3. Vortex the capped tube for 30 seconds.
1. Intended Use 4. Place the capped tube in over 90°C (194˚F) water for 10 minutes.
Rapid Test ProⅡ is intended for a quick detection of protein from allergenic ingredients in samples.
5. Place the tube under running water to cool down to ambient temperature.
6. Vortex for 30 seconds.
2. Features 7. Stand the tube for a few minutes to let the sample settle down, and then collect the supernatant
A qualitative test in lateral flow immunoassay format for visual detection
as Sample Extract.
Available for food products, swabs and Clean-in-place rinse water
NOTE: Centrifuge and/or filter with filter paper as required.
Provide test results in 15 minutes (Sample preparation time: within 30 minutes)
Improved recovery of proteins from both unprocessed and processed food products by the invented
extraction solution1-4) 2 3 4
Sensitivity of each kit is shown in Table1,2 Extraction Solution
(19 mL)
Table1: Sensitivity of total protein in Rapid Test ProⅡ for Egg, Casein, Buckwheat,
Peanut and Soya (by Recommended Extraction Method)
Test sample*
Kit Calculated as Food Sample (1.0 g) in
Swab a polypropylene
Rinse water
tube (L)
Rapid Test ProⅡ for Egg Egg total protein Vortex for 30 seconds Over 90°C for 10 minutes
Rapid Test ProⅡ for Casein Milk total protein
1 μg/swab
Buckwheat total 5 μg/g (μg/mL) 5 6
Rapid Test ProⅡ for Buckwheat (When adding 4 mL of
protein (5 ppm)
Extraction Solution)
Rapid Test ProⅡ for Peanut Peanut total protein
Rapid Test ProⅡ for Soya Soya total protein
*Sensitivity in Test Solution: 25 ng/mL
Table2: Sensitivity of Gluten in Rapid Test ProⅡ for Gluten
(by Recommended Extraction Method)
Test sample** Cool down Vortex for 30 seconds
Kit Calculated as Food
Swab
Rinse water A-2. For food sample【Simplified Extraction Method】
0.8 μg/swab In some samples, the heating (procedures 4-6) may be omitted from the Recommended Extraction
4 μg/g (μg/mL)
Rapid Test ProⅡ for Gluten Gluten* (When adding 4 mL of Method. However, the Simplified Extraction method may cause invalid, false-negative or false-positive
(4 ppm)
Extraction Solution) results. For further information please contact us.
*Calculated using a conversion factor: 0.85 ng gluten per ng total whole wheat protein5-8)
**Sensitivity in Test Solution: 20 ng/mL. B-1. For swab test sample【Recommended Extraction Method】
1. Thoroughly wipe off a specified surface area with a Cotton swab moistened with tap water.
3. Kit Components 2. Place the swab into a polypropylene tube (L) and add 4 mL of Extraction Solution.
(Sensitivity: 1 μg (0.8 μg for gluten) on the swab in the case of adding 4 mL, see Table1, 2)
Component Amount
NOTE: The sensitivity can be adjusted by changing the volume of Extraction Solution.
Rapid Test ProⅡ
When adding 19 mL of Extraction Solution, the sensitivity is 5 μg (4 μg for gluten) on the swab.
Extraction Solution* 10 packs (19 mL/pack) 3. Cap the tube tightly and vortex it for 30 seconds.
Diluent** 1 bottle (12 mL) 4. Place the capped tube in over 90°C (194˚F) water for 10 minutes.
Test stick 10 packs (1 stick/pack) 5. Place the tube under running water to cool down to ambient temperature.
Pipette (L) 10 6. Vortex for 30 seconds. The resulting solution is referred to as Sample Extract.
Pipette (S) 10 NOTE: Filter it with filter paper as required.
Polypropylene tube (L), 50 mL volume 10
Polypropylene tube (S), 1.5 mL volume 10
1 2 3
Cotton swab 10 packs Add 4 mL of Extraction Solution
Paper tube rack 1 using a scale
on a polypropylene tube(L)
* Extraction Solutions for all test kits are identical. Extraction Solution may contain precipitates
under refrigeration that should dissolve upon warming at 30−37˚C (86−99˚F).
** Diluents for Egg kit, Casein kit and Gluten kit are identical. Diluents for Buckwheat kit and Peanut
are identical. Diluent for Soya kit is only used for Soya.
The swab
Sample application slot into
Test window Wipe off a polypropylene
4 mL→ tube (L) Vortex for 30 seconds
Confirmation slot
4 5 6
Test stick
1 mL 2 mL
↓ ↓
Pipette (L)
Over 90°C for 10 minutes Cool down Vortex for 30 seconds
100 μL 200 μL 300 μL 4 mL→
↓ ↓ ↓
B-2. For swab test sample【Simplified Extraction Method】
Polypropylene tube (L) In some samples, the heating (procedures 4-6) may be omitted from the Recommended Extraction
Pipette (S)
50 mL volume Method. However, the Simplified Extraction Method may cause invalid, false-negative or false-positive
results. For further information please contact us.
C-1. For rinse water sample【Recommended Extraction Method】 8. Assessment criteria
1. Put 1.0 g (mL) of the sample in a polypropylene tube (L) and add 1 pack of Extraction Solution. Precautions
2. Vortex the capped tube for 30 seconds. Test results must be judged just in 15 minutes even though there appears red-purple color in a test
3. Place the capped tube in over 90°C (194˚F) water for 10 minutes. window during the reaction.
4. Place the tube under running water to cool down to ambient temperature.
5. Vortex for 30 seconds. The resulting solution is referred to as Sample Extract. Sample application slot
Test window
Confirmation window
1 2 3
Extraction Solution
(19 mL) A. Positive
B. Negative
Sample (1.0 g) in
a polypropylene C. Invalid
tube (L)
Vortex for 30 seconds Over 90°C for 10 minutes
A) Positive: A red-purple line in a test window together with red color in a confirmation window.
4 5 B) Negative: No line in a test window together with a red color in a confirmation window.
C) Invalid: No color in a confirmation window.
NOTE: If no color in a confirmation window, retest with a new stick. Negative results will occur if Test
Solution contains protein less than detectable levels. False-negative results may occur if Test
Solution contains protein at relatively high concentrations, in that case, retest with a diluted Test
Solution (see 6. Preparation of Test Solution).
Cool down Vortex for 30 seconds 9. Warnings
1. Do not combine reagents from different lots.
2. Store the kit at 2−8˚C (35−46˚F), and DO NOT FREEZE.
C-2. For rinse water sample【Simplified Extraction Method】 3. Do not use the kit after the expiration date indicated on the box.
In some samples, the heating (procedures 3-5) may be omitted from the Recommended Extraction
Method. However, the Simplified Extraction method may cause invalid, false-negative or false-positive 10. References
results. For further information please contact us. 1. Patent No.: JP 5133663
2. Patent No.: AU 2008330507
6. Preparation of Test Solution 3. Patent No.: US 8,859,212
1. Dispense 900 µL of Diluent with a pipette (L) into a polypropylene tube (S). 4. Patent No.: EP 2224239 (AT, BE, DE, ES, FR, GB, IT, NL, CH)
2. Add 100 µL of Sample Extract with a pipette (S) to the polypropylene tube (S) containing 900 µL 5. T. B. Osborne (1924), The Vegetable Proteins, Longmans and Co., London, UK.
of Diluent and mix well. The resulting solution is referred to as Test Solution. 6. John Holme (1966), “A review of Wheat Flour Proteins and their functional properties”, The Bakers
Digest, Vol.40, pp.38−42.
NOTE: For further dilution, dilute the Sample Extract with Extraction Solution appropriately, and 7. S. Shibata and T. Nakae (Eds.) (1990), Komugikoseihinno chisiki [Knowledge of wheat products],
then dilute it 10-fold with Diluent. Saiwai Shobo, Tokyo, Japan.
8. Nihon mugirui kenkyukai (Ed.) (1964), Komugiko-sono genryou to kakouhin [Wheat-material and
processing], Nihon mugirui kennkyukai, Tokyo, Japan.
2-1 Pipette (S)
11. Warranties
Morinaga Institute of Biological Science, Inc. makes no warranty of any kind, either expressed or
100 µL implied, except that the materials from which its products are made are of standard quality. Buyer
assumes all risk and liability resulting from the use of this product. There is no warranty of
1 merchantability of the product, or of the fitness of the product for any purpose. Morinaga Institute of
Pipette (L) Biological Science, Inc. agrees to replace any defective product, but expressly disclaims liability for
damages, including special or consequential damage, or expenses arising directly or indirectly from the
900 µL
900 µL Food or rinse water use of this product.
test sample
12. Appendix: Test Flow Chart
2-2 Pipette (S)
A. Food sample
B. Swab test sample
C. Rinse water sample
Diluent 100 µL
Mix and homogenize a sample
Swab a specified area
(for A. Food sample)
▼ ▼
Put 1.0 g (mL) of the homogenized
Place the swab into the tube and
sample in a tube and add 1 pack of
add 4 mL of Extraction Solution
Extraction Solution
Swab test sample
▼ ▼
Vortex for 30 seconds
5. Sample Extraction
▼ ▼
Place the tube in over 90˚C (194˚F) water for 10 minutes
7. Assay procedures
and cool down*
Precautions
▼ ▼
Prior to use, place a Test stick to 20−30˚C (68−86˚F) and open the package just before use.
Vortex for 30 seconds*
A low-temperature test stick may not work properly.
▼ ▼
Do not touch a sample application slot and a test window of a Test stick. Let the sample settle down and collect the supernatant
as Sample Extract
1. Place a Test stick horizontally and add 200 µL of Test Solution to the sample application slot. ▼ ▼
2. Incubate 15 minutes at room temperature. 6. Preparation of Add 100 µL of Sample Extract to 900 µL of Diluent and mix well
3. Interpret the results in a test window according to 8. Assessment criteria described below. Test Solution as Test Solution
▼ ▼
7. Assay procedures Add 200 µL of Test Solution to a Test stick
1 Pipette (S) ▼ ▼
8. Assessment criteria Check results in 15 minutes
200 µL *Simplified Extraction Method may be applicable to some samples; the operations such as heating and
vortexing can be omitted. However, the method may cause false-negative or false-positive results. For
further information please contact us (http://www.miobs-e.com).
Test stick
Test solution
