BIOO FOOD AND FEED SAFETY

Oxytetracycline ELISA Test Kit Manual

Catalog #: 1081
Reference #: FO1081-01A

This kit is manufactured to the international quality standard ISO 9001:2008.

ISO CI#: SARA-2009-CA-0114-02-A

BIOO Scientific Corp. 2014

TABLE OF CONTENTS
GENERAL INFORMATION ...................................................................... 1
Product Description .................................................................................. 1
Procedure Overview.................................................................................. 1
Kit Contents, Storage and Shelf Life ......................................................... 2
Sensitivity (Detection Limit) ..................................................................... 2
Specificity (Cross-Reactivity).................................................................... 2
Required Materials Not Provided With the Kit ......................................... 3
Warnings and Precautions ........................................................................ 3
SAMPLE PREPARATION .......................................................................... 4
Honey ........................................................................................................ 4
Meat/Fish/Shrimp ..................................................................................... 4
Milk ........................................................................................................... 5
Milk Powder .............................................................................................. 5
Serum ........................................................................................................ 5
Urine ......................................................................................................... 6
OXYTETRACYCLINE ELISA TEST KIT PROTOCOL ....................... 6
Reagent Preparation ................................................................................. 6
ELISA Testing Protocol ............................................................................. 7
Oxytetracycline Concentration Calculations ........................................... 8
TROUBLESHOOTING ............................................................................... 9
No Color Development or No Signals with Standards ............................. 9
Low Optical Density (OD) Readings ........................................................ 9
High Background or High Optical Density (OD) Readings..................... 9
High Intra-Plate Variance ...................................................................... 10
High Inter-Plate Variance ....................................................................... 10
One or More of the Standard Curve Points Are Out of Range ............... 10
The MaxSignal Oxytetracycline ELISA Test Kit is intended for laboratory use only, unless otherwise indicated. This
product is NOT for clinical diagnostic use. MaxSignal is a registered trademark of Bioo Scientific Corporation (BIOO).

MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

GENERAL INFORMATION
GENERAL INFORMATION

Product Description
The MaxSignal Oxytetracycline ELISA Test Kit is a competitive enzyme immunoassay for the
quantitative analysis of oxytetracycline in honey, milk, urine, fish, shrimp and meat.
Oxytetracycline is a broad spectrum antibiotic which is frequently employed in animal
production for its excellent antibacterial and pharmacokinetic properties.
The MaxSignal Oxytetracycline ELISA Test Kit enables international and government
regulatory agencies, food manufacturers and processors, as well as quality assurance
organizations, to detect oxytetracycline in honey, milk, fish, shrimp or meat and to satisfy
customer concerns about food safety. The unique features of the kit are:

High recovery (>80%), rapid (less than 30 minutes), and cost-effective extraction methods.
High sensitivity (0.15 ng/g or ppb) and low detection limit.
A quick ELISA assay (less than 2 hours regardless of number of samples).
High reproducibility.

Procedure Overview
The method is based on a competitive colorimetric ELISA assay. The drug of interest has been
coated in the plate wells. During the analysis, sample is added along with the primary antibody
specific for the target drug. If the target is present in the sample, it will compete for the antibody,
thereby preventing the antibody from binding to the drug attached to the well. The secondary
antibody, tagged with a peroxidase enzyme, targets the primary antibody that is complexed to
the drug coated on the plate wells. The resulting color intensity, after addition of substrate, has
an inverse relationship with the target concentration in the sample.

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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

Kit Contents, Storage and Shelf Life

The MaxSignal Oxytetracycline ELISA Test Kit has the capacity for 96 determinations or
testing of 42 samples in duplicate (assuming 12 wells for standards). Return any unused
microwells to the foil bag and reseal them with the desiccant provided in the original package.
Store the kit at 2-8C *. The shelf life is 12 months when the kit is properly stored.
Kit Contents
Oxytetracycline-coated Plate
Oxytetracycline Standard Stock
(450 ng Powder)
Empty Vials for Standards:
Negative control (white CAP tube)
0.15 ng/mL (yellow CAP tube)
0.375 ng/mL (orange CAP tube)
0.75 ng/mL (pink CAP tube)
1.5 ng/mL (purple CAP tube)
4.5 ng/mL (blue CAP tube)
Oxytetracycline Spiking Stock
(2000 ng Powder)
Oxytetracycline Antibody #1
100X HRP-Conjugated Antibody #2
Antibody #2 Diluent **
20X Wash Solution **
Stop Buffer **
TMB Substrate**
10X Tissue Extraction Buffer (Optional)
TCA Buffer Additive (Optional)
TCA Balance Buffer (Optional)
Standard Diluent
10X OXYTET Balance Buffer (Optional)

Amount
1 x 96-well Plate (8 wells x 12 strips)
3
1
1
1
1
1
1
1
12 mL
0.3 mL
20 mL
28 mL
14 mL
12 mL
25 mL
5 mL
28 mL
28 mL
10 mL

Storage
2-8C
-20C

2-8C

-20C
2-8C *
2-8C *
2-8C
2-8C
2-8C
2-8C
2-8C
2-8C
2-8C
2-8C
2-8C

* If you are not planning to use the kit for over 1 month, storing Oxytetracycline Antibody
#1 and 100X HRP-Conjugated Antibody #2 at -20C or in a freezer is recommended.
** Components with the same BIOO part number within their expiration dates are
interchangeable among BIOO kits.

Sensitivity (Detection Limit)

Sample Type
Honey
Meat/Fish/Shrimp
Milk
Milk powder
Serum
Urine

Detection Limit (ng/g or ppb)

4
3
4
40
6
6

Specificity (Cross-Reactivity)
Analytes
Oxytetracycline
Chlortetracycline
Minocycline

Cross-Reactivity (%)
100.0
108
79

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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

Demeclocycline 37
Tetracycline 124
Doxycycline 62

Required Materials Not Provided With the Kit

Microtiter plate reader (450 nm)

Vortex mixer, (e.g. Genie Vortex mixer from VWR)
10, 20, 100 and 1000 L pipettes
Multi-channel pipette: 50-300 L (Optional)
10 mM PBS buffer (1X PBS): 0.24 g KH2PO4 + 1.44 g Na2HPO4 + 8 g NaCl, + 0.2 g KCl,
adjust pH to 7.4 with NaOH, fill up to 1000 mL with distilled water

Warnings and Precautions

BIOO strongly recommends that you read the following warnings and precautions to
ensure your full awareness of ELISA techniques and other details you should pay close
attention to when running the assays. More information can also be found in
Troubleshooting section. Periodically, optimizations and revisions are made to the kit and
manual. Therefore, it is important to follow the protocol coming with the kit. If you need
further assistance, you may contact your local distributor or BIOO at
techsupport@biooscientific.com.

The standards contain oxytetracycline. Handle with particular care.

Do not use the kit past the expiration date.
Do not mix reagents from different kits or lots except for components with the same part
Nos within their expiration dates. ANTIBODIES AND PLATES ARE KIT-AND
LOT-SPECIFIC.
Try to maintain a laboratory temperature of 2025C (6877F). Avoid running assays
under or near air vents, as this may cause excessive cooling, heating and/or evaporation.
Also, do not run assays in direct sunlight, as this may cause excessive heat and
evaporation. Cold bench tops should be avoided by placing several layers of paper towel
or some other insulation material under the assay plates during incubation.
Make sure you are using only distilled or deionized water since water quality is very
important.
When pipetting samples or reagents into an empty microtiter plate, place the pipette tips in
the lower corner of the well, making contact with the plastic.
Incubations of assay plates should be timed as precisely as possible. Be consistent when
adding standards to the assay plate. Add your standards first and then your samples.
Add standards to plate only in the order from low concentration to high concentration as
this will minimize the risk of compromising the standard curve.
Always refrigerate plates in sealed bags with a desiccant to maintain stability. Prevent
condensation from forming on plates by allowing them equilibrate to room temperature (20
25C / 68 77F) while in the packaging.
BIOO makes no warranty of any kind, either expressed or implied, except that the materials from which its
products are made are of standard quality. There is no warranty of merchantability of this product, or of the
fitness of the product for any purpose. BIOO shall not be liable for any damages, including special or
consequential damage, or expense arising directly or indirectly from the use of this product.

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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

SAMPLE PREPARATION
SAMPLE PREPARATION
Be sure samples are properly stored. In general, samples should be refrigerated at 2-4C for
no more than 1-2 days. Freeze samples to a minimum of -20C if they need to be stored for a
longer period. Frozen samples can be thawed at room temps (20 25C / 68 77F) or in a
refrigerator before use.
1. Preparation of 1X Tissue Extraction Buffer
Mix 1 volume of 10X Tissue Extraction Buffer with 9 volumes of distilled water.

2. Preparation of 10% TCA Extraction Buffer

Weigh 20 g of TCA (trichloroacetic acid), add 175 mL of distilled water and 5 mL of TCA Buffer
Additive, mix well.

3. Preparation of 1X OXYTET Balance Buffer

Mix 1 volume of 10X OXYTET Balance Buffer with 9 volumes of distilled water.

4. Preparation of Oxytetracycline Spiking Standard

The oxytetracycline spiking is provided as 2000 ng powder stock. Take out the stock
powder, add 1.0 mL of Standard Diluent. Vortex vigorously for 2 minutes, then leave this
solution at room temperature at dark for 15 minutes. Then vortex vigorously again for 1
minute and leave it at room temperature at dark for another 15 minutes. Vortex the solution
for about 30 seconds for a third time, and then transfer this 2000 ppb stock solution to a
dark plastical tube and store it at -20C for any future use.

Honey
1. Weigh out 0.5 g of honey in a screw-top glass vial (50 mL)
2. Add 12 mL 10 mM PBS buffer, pH 7.4.
3. Put the solution in an ultrasonic bath for 5 min and vortex for 2 min; alternatively the
sample can be mix vigorously for 10 minutes in a multi-tube vortexer or shaker.
4. Use 75 L of the sample for the assay.

Note: Dilution factor: 25.

Meat/Fish/Shrimp
Method I
1. Add 2 mL of 1X Tissue Extraction Buffer to 1 g of homogenized meat, fish or shrimp, vortex
for 15 minutes in a multi-tube vortexer or shake 30 minutes on a shaker.
2. Centrifuge for 10 minutes at 4,000 x g.
3. Transfer 300 L of the supernatant to a new tube containing 700 L of 1X OXYTET
Balance Buffer. Vortex for 30 seconds.
4. Use 75 L per well in the assay.
Note: Dilution factor: 10 (detection range: 1.5-45 ppb). The dilution factor can
be changed to 60 ((detection range: 9-270 ppb)if Step 3 is changed to Transfer
30 L of supernatant to a new tube containing 570 L of 1X OXYTET Balance
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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

Buffer. Vortex for 30 seconds. The dilution factor can be changed to 80

((detection range: 12-360 ppb)if Step 3 is changed to Transfer 30 L of
supernatant to a new tube containing 770 L of 1X OXYTET Balance Buffer.
Vortex for 30 seconds.
Method II (For samples with low recovery rate extracted by Method I)
1. Add 2 mL of 10% TCA Extraction Buffer and 2 mL of hexane to 1 g of homogenized meat,
fish or shrimp sample, vortex for 15 minutes in a multi-tube vortexer or manually vortex 3
minutes at maximum speed.
2. Centrifuge for 10 minutes at 4,000 x g.
3. Transfer 240 L of the bottom aqueous layer to a new tube containing 340 L of TCA
Balance Buffer.
Note: Be careful to avoid any hexane layer or interphase that has formed. Do
this by passing the pipette tip slowly through these layers and only removing
from the aqueous layer.
4. Leave the tube open and allow the solution to evolve gas while gently swirling the tube.
5. After 5 minutes, add 220 L of Standard Diluent and vortex for 1 minute.
Note: If gas bubbles are still forming, leave the tube open for another 2 5
minutes while swirling in order to avoid any spilling of the samples.
6. Use 75 L per well in the assay.
Note: Dilution factor: 10 (detection range: 1.5-45 ppb).

Milk
1. For regular milk with fat, centrifuge 1.5 mL of the cold milk sample (~10C / 50F ) at

10000 rpm for 10 minutes. Discard the upper fat layer. ( For fat-free milk, skip this step).

2. Take 100 L of the milk sample and add 0.9 mL of 10 mM PBS buffer, pH 7.4. Vortex the

tube for 30 seconds.

3. Use 75 L of the sample for the assay.

Note: Dilution factor: 10.

Milk Powder
1. Weigh out 1 g of milk powder in a centrifugal glass vial, add 9 mL of distilled water and

dissolve by shaking.
2. Centrifuge at 10000 rpm for 10 minutes, discard the upper lipid layer.
3. Take 100 L of the milk sample and add 0.9 mL of 10 mM PBS buffer, pH 7.4. Vortex the
tube for 30 seconds.
4. Use 75 L of the sample for the assay.
Note: Dilution factor: 90

Serum
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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

1.
2.
3.
4.

Centrifuge 0.2 mL of the serum sample at 4,000 rpm for 5 minutes.

Take 0.1 mL of supernatant, add 3.9 mL of 10 mM PBS buffer, pH 7.4.
Vortex for 1 minute.
Use 75 L of the sample for the assay.
Note: Dilution factor: 40.

Urine
5.
6.
7.
8.

Centrifuge 1.5 mL of the urine sample at 4,000 rpm for 5 minutes.

Take 0.5 mL of supernatant, add 19.5 mL of 10 mM PBS buffer, pH 7.4.
Vortex for 1 minute.
Use 75 L of the sample for the assay.
Note: Dilution factor: 40.
Preparation protocols for samples other than above can be made available upon request.
Please contact your local distributor or write us at foodfeedsafety@biooscientific.com.

OXYTETRACYCLINE
ELISA TEST
KITTEST
PROTOCOL
OXYTETRACYCLINE
ELISA
KIT PROTOCOL

Reagent Preparation
IMPORTANT: All reagents should be brought up to room temperature before use (1 2
hours at 20 25C / 68 77F); Make sure you read Warnings and Precautions section.
Solutions should be prepared just prior to ELISA test. All reagents should be mixed by gently
inverting or swirling prior to use. Prepare volumes that are needed for the number of wells
being run. Do not return the reagents to the original stock tubes/bottles. Using disposable
reservoirs when handling reagents can minimize the risk of contamination and is
recommended.
1. Preparation of Oxytetracycline Work Standards

The oxytetracycline standard is provided as 450 ng stock. Add 1.5 mL of Standard Diluent
to the stock vial and mix by vortexing for 1 minute and leave at room temperature for at
least 30 minutes to obtain 300 ppb stock vial. Mix the stock vial again by shaking up and
down 10 times before use. To make work standards, serially dilute the 300 ppb standard
stock vial with Standard Diluent, mix well by vortexing each work standard tube for 30
seconds.
Work Standards Oxytetracycline
Source
4.5 ppb vial 300 ppb stock vial
1.5 ppb vial 4.5 ppb vial
0.75ppb vial 1.5 ppb vial
0.375 ppb vial 0.75 ppb vial
0.15 ppb vial 0.375 ppb vial
Negative Control vial N/A

Volume of Source
Oxytetracycline
30 L
500 L
500 L
500 L
500 L
0 L

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Volume of Standard
Diluent
1970 L
1000 L
500 L
500 L
750L
500 L
6

MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

Note: The 300 ppb stock vial and the work standards can be used within one week if
stored at 4C. The empty standard vials can be stored at 4C and re-used for the next
standard preparation.
2. Preparation of 1X Wash Solution

Mix 1 volume of the 20X Wash Solution with 19 volumes of distilled water.
3. Preparation of 1X HRP-Conjugated Antibody #2

Mix 1 volume of the 100X HRP-Conjugated Antibody #2 with 99 volumes of Ab#2 Diluent.
4. Some special notes about this kit

Please take out the whole kit, put it at room temperature for at least two hours before the
assay; Try to avoid light as much as you can during sample preparation and assay running;
Try to wash the plate a little bit harder shake the plate with 300 L of 1X Wash Buffer for
15 seconds before dumping the buffer and dry the plate, do it again for two more times and
dry the plate well in the third time; Pipette the reagents and samples very accurately
(especially the samples) even though you have to slow down pipetting.

ELISA Testing Protocol

Label the individual strips that will be used and aliquot reagents as the following example:
Component
Oxytetracycline Antibody #1
1X HRP-Conjugated Antibody #2
1X Wash Solution
Stop Buffer
TMB Substrate

Volume per Reaction

100 L
150 L
2.5 mL
100 L
100 L

24 Reactions
2.4 mL
3.6 mL
60 mL
2.4 mL
2.4 mL

1. Add 75 L of each Oxytetracycline Standard (Negative Control, 0.15, 0.375, 0.75. 1.5, 4.5

ppb) in duplicate into different wells ( Add standards to plate only in the order from
low concentration to high concentration).

2. Add 75 L of each sample in duplicate into different sample wells.

3. Add 100 L of Antibody #1 and mix well by gently rocking the plate manually for 1 minute.
4. Incubate the plate for 50 minutes at room temperature (20 25C / 68 77F).
5. Wash the plate 3 times with 250 L of 1X Wash Solution. After the last wash, invert the

plate and gently tap the plate dry on paper towels ( Perform the next step immediately
after plate washings. Do not allow the plate to air dry between working steps).

6. Add 150 L of 1X Antibody #2 solution. Incubate the plate for 20 minutes at room

temperature (20 25C / 68 77F) ( Avoid direct sunlight and cold bench tops
during the incubation. Covering the microtiter plate while incubating is
recommended).
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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

7. Wash the plate 3 times with 250 of L 1X Wash Solution. After the last wash, invert the

plate and gently tap the plate dry on paper towels ( Perform the next step immediately
after plate washings. Do not allow the plate to air dry between working steps).

8. Add 100 L of TMB Substrate to each well. Time the reaction immediately after adding the

substrate. Mix the solution by gently rocking the plate manually for 1 minute while
incubating ( Do not put any substrate back to the original container to avoid
any potential contamination. Covering the microtiter plate while incubating is
recommended).

9. After incubating for 15 minutes at room temperature (20 25C / 68 77F), add 100

L of Stop Buffer to stop the enzyme reaction.

10. Read the plate as soon as possible following the addition of Stop Buffer on a plate reader

with 450 nm wavelength ( Before reading, use a lint-free wipe on the bottom of the
plate to ensure no moisture or fingerprints interfere with the readings).

Oxytetracycline Concentration Calculations

A standard curve can be constructed by plotting the mean relative absorbance (%) obtained
from each reference standard against its concentration in ng/mL on a logarithmic curve.
Relative absorbance (%) =

absorbance standard (or sample) x 100

absorbance zero standard

Use the mean relative absorbance values for each sample to determine the corresponding
concentration of the tested drug in ng/mL from the standard curve. A special program with
Excel functionality, the MaxSignal ELISA Detection Analysis System, is available upon
request to evaluate the MaxSignal ELISA test results. Please contact your local distributor or
techsupport@biooscientific.com for further information. The following figure is a typical
oxytetracycline standard curve.

Relative Absorbance (%)

O xyte tracycline Standard Curve

90
70
50
30
10
0.01

0.10

1.00

10.00

100.00

Standard ng/mL (LN)

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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

TROUBLESHOOTING TROUBLESHOOTING

No Color Development or No Signals with Standards

Possible Causes

Recommended Action

Reagents were used in the wrong

order or a step was skipped.

Follow the protocol carefully and repeat the assay.

Wrong antibodies were used.

Substrate has deteriorated.

Make sure that the antibodies used are the ones that came with the kit. All
antibodies are kit- and lot-specific.
Use a new set of BIOO Substrate.

Low Optical Density (OD) Readings

Possible Causes
Reagents were expired or mixed
with a different lot number.
Wash solution was prepared
incorrectly.
Too many wash cycles were used.
Incubation times were too short.
Lab temperature was too low.
Reagents and plates were too cold.
Reader was at wrong wavelength,
or reader was malfunctioning.
Excessive kit stress has occurred.
Assay plates were compromised.

Recommended Action
Verify the expiration dates and lot numbers.
Use the wash solution for the kit and make sure that it is prepared correctly.
Make sure to use the number of washes per the protocol instruction.
Time each plate separately to ensure accurate incubation times, follow protocol.
Maintain the lab room temperature within 2025C (6877F). Do not run
assays under air conditioning vents or near cold windows.
Make sure plates and reagents are brought up to room temperature. Keep the
kit components out of the kit box for at least 1 hour before starting the assay.
Make sure the wavelength is 450 nm for the assay and read the plate again.
Verify reader calibration and lamp alignment.
Check records to see how many times the kit has cycled from the refrigerator.
Check to see if the kit was left at extreme temperatures for too long.
Always refrigerate plates in sealed bags with a desiccant to maintain stability.
Prevent condensation from forming on plates by allowing them equilibrate to
room temperature (20 25C / 68 77F) while in the packaging.

High Background or High Optical Density (OD) Readings

Possible Causes

Recommended Action

Poor quality water was used in

wash solution.
Substrate solution has deteriorated.

If water quality is questionable, try substituting an alternate distilled water

source to prepare the wash solution.
Make sure the substrate is colorless prior to addition to the plate.
Use the number of washes per the protocol instruction. Make sure that at least
250 L of wash solution is dispensed per well per wash. Verify the performance
of the washer system; have the system repaired if any ports drip, dispense or
aspirate poorly.

There was insufficient washing or

poor washer performance.
Reader was malfunctioning or not
blanked properly. This is a high
possibility if the OD readings were
high and the color was light.

Verify the readers performance using a calibration plate and check the lamp
alignment. Verify the blanking procedure, if applicable, and reblank.

Lab temperature was too high.

Maintain the room temperature within 2025C (6877F). Avoid running

assays near heat sources or in direct sunlight.

Reagents were intermixed,

contaminated or prepared
incorrectly.

Ensure that the correct reagents were used, that working solutions were
prepared correctly and that contamination has not occurred.

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MaxSignal Oxytetracycline ELISA Test Kit Manual FO1081-01A

High Intra-Plate Variance

Possible Causes

Recommended Action

Inconsistent time was taken when

adding standards, reagents or
samples to the assay plate.
Multichannel pipette was not
functioning properly.
There was inconsistent washing or
washer system malfunctioning.

Make sure all materials are set up and ready to use. Use a multichannel pipette
to add reagents to multiple wells whenever possible. Do not interrupt while
adding standards, reagents and samples.
Verify pipette calibration and check that tips are on tight. Be sure all channels of
the pipette draw and dispense equal volumes.
Check performance of the wash system. Have the system repaired if any ports
drip or dispense/aspirate poorly.

High Inter-Plate Variance

Possible Causes

Recommended Action

Inconsistent incubation times

occurred from plate to plate.

Time each plate separately to ensure consistent incubation times.

Inconsistent washing occurred from

plate to plate.
Pipette was working improperly.
Kit plates, reagents, standards and
samples were at different
temperatures.
Reagents used were intermixed
from different kit lots, or the kits
were of different expiration dates.

Make sure to use the number of washes per the protocol instruction. Verify
performance of the wash system and have the system repaired if any ports drip
or dispense/ aspirate poorly.
Check the pipette calibration. Verify that pipette tips are on tight before use and
that all channels draw and dispense equal volumes.
Make sure to allow sufficient time for kit plates, reagents, standards and
samples come to room temperature (20 25C / 68 77F). Larger volumes
will require longer equilibration time. If using a water bath to hasten
equilibration, make sure it is maintained at room temperature; do not use a
warm water bath to warm reagents, samples and kit standards.
Carefully label each reagent to make sure the reagents are not intermixed. Kits
with different expiration dates might generate different range of OD readings,
however, the relative absorbance values may very well be comparable. In
general, a value of less than 0.6 in zero standard reading may indicate certain
degrees of deterioration of reagents.

One or More of the Standard Curve Points Are Out of Range

Possible Causes

Recommended Action

Standards were added in wrong

order or recorded in wrong position.
Standards were contaminated or
intermixed with other standards.
There was inconsistent washing or
washer system malfunctioning.

Follow the protocol and re-run the assay. Make sure the standards are applied
and recorded correctly.
Use a new set of standards. Add standards to plate only in the order from low
concentration to high concentration.
Perform washing consistently. Check performance of the wash system. Have
the system repaired if any ports drip or dispense/aspirate poorly.
Make sure all materials are set up and ready to use. Add standards to plate only
in the order from low concentration to high concentration at undisrupted pace.
Use a multichannel pipette to add reagents to multiple wells simultaneously.
Verify pipette calibration and check that tips are on tight. Be sure all channels of
the pipette draw and dispense equal volumes.

Inconsistent time was taken to add

standards and reagents to plate.
Multichannel pipette was not
functioning properly.

Bioo Scientific Corporation

7050 Burleson Road
Austin, TX 78744 USA
Tel: 1.888.208.2246
Fax: (512) 707-8122

Made in USA
BIOO Food & Feed Safety Products
info@biooscientific.com
foodfeedsafety@biooscientific.com
www.biooscientific.com

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