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Our mission is to provide you with a higher level of technical support. We encourage you to use our site to find valuable information to help you troubleshoot common user problems, review frequently asked questions, access technical notes (Technical Library), or send us a suggestion as to how we can improve upon our service.

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FAQ

You can request a CoA by filling out the form in our Technical Library.

On each product page you can find the latest version of the DfU in English. If you need other versions of DfU, please send us your request by filling out the form in our Technical Library.

On each product page you can find the latest version of the SDS in English. If you need other versions of the SDS, please send us your request by filling out the form in our Technical Library.

You can find all technical notes in our Technical Library.

Plate sealers should not be used if not specifically stated in the Directions for Use. Sealing the plate may have a negative effect on the results if the assay has been designed and developed without the use of plate sealers. If the liquid in the wells is splashing during shaking of the plates, you need to check the settings of your shaker.

Plate sealers are included in all Mercodia kits where they should be used according to the Directions for Use, such as Mercodia Glucagon ELISA and Total GIP NL-ELISA.

The expiry date is stated on the label of the kit and in the Certificate of Analyis (CoA). You can request a CoA by filling out the form in our Technical Library.

The sample reaction in Mercodia assays might be affected by pH and salt concentration. To ensure a proper reaction environment, sample buffer or Calibrator 0 included in the Mercodia ELISA kit should be used to dilute serum/plasma samples, antigen extraction or antigen in growth medium. Further information about dilution of samples is stated in the Directions for Use of each product. PBS or other salt solution is not recommended for sample dilution.

For several of the Mercodia ELISAs, which include all insulin assays, all proinsulin assays and Rat C-peptide ELISA, customers may separately order Diabetes Sample Buffer (10-1195-01) that can be used for diluting samples.

The validated sample types are stated in the Directions for Use (DfU).

For sample types other than those recommended in the DfU, a test to evaluate sample type and matrix compatibility can be performed to ensure reliable results. See technical note TN34-0123 in our Technical Library.

Calibrator 0 should not be used for blank subtraction, but merely be regarded as a negative control.

As stated in the Directions for Use of each product, a Mercodia ELISA is validated using Magellan software (Tecan) for absorbance-based ELISA, or MARS (BMG Labtech). You should be able to use similar sofwares to do your calculations.

Alternatively, it is also possible to perform curve-fitting, including both Five Parameter Logistic (5PL) and cubic spline, as well as data interpolation in commercial statistical software such as GraphPad Prism, or websites such as MyAssays.

When using website such as MyAssays, add the negative control (calibrator 0) equal to zero for the blank in the plate layout. The reason is that a blank is obligatory to add in the plate layout for the website, and Mercodia’s ELISA is not validated with blank reduction. For further information regarding how to use MyAssays, please see technical note TN34-0167 in our Technical Library.

You should never extrapolate the curve to values outside of the range of the calibrators. Instead assign the concentrations <Cal 1 or >Cal5.

You are always welcome to reach out to us at support@mercodia.com but you can also find helpful troubleshooting tips in the Troubleshooting section below.

We strive to get back to you with an order confirmation within 24 hours of receiving your order. It could take longer depending on the current workload but feel free to send us a new email after 24 hours to double-check.

Within 2 days from the receipt of your order confirmation is our general rule. However, we do not ship within Europe on Fridays.

Yes, the products will meet specifications even if the package is delayed for several weeks. Our products are shipped in ambient temperature worldwide and have been stress and stability tested to make sure this is safe. Please see our Stability Statement here.

The plates used in the Mercodia assays are flat bottomed Nunc 96 plates produced by Thermo Fischer.

TROUBLESHOOTING

Sources of error	Cause	Background/Suggestions
Contamination of substrate	Substrate was contaminated by enzyme conjugate solution.	Always use clean pipette tips, beakers etc. in order to avoid contamination. Rinsing with water is not enough. Containers and pipette tips used for enzyme conjugate solution should be kept separate from the substrate.
Contamination by samples or calibrators	Assay- or Sample Buffer was contaminated by samples or calibrators.	Always use clean pipette tips, beakers etc. in order to avoid contamination.
Washing procedure	Insufficient washing before substrate incubation.	Plate washer should be checked and served periodically. When washing manually use a laboratory wash bottle, see Tech note 34-0106 Instruction for manual washing (Technical Library).
Incorrect dilution of enzyme conjugate solution	Too much enzyme conjugate stock solution has been added to the Enzyme Conjugate Buffer.	Dilute enzyme conjugate solution according to Directions for Use.

Sources of error	Cause	Background/Suggestions
Stop Solution	Substrate TMB was contaminated by stop solution prior to addition to wells. The assay procedure has been done incorrectly.	Avoid contaminating Substrate TMB with Stop Solution by labeling any containers used. Always make sure that the stop solution is added after the Substrate TMB. The Stop Solution causes the enzyme-substrate reaction to reach end-point.
Enzyme conjugate solution	No enzyme conjugate stock solution was added to the Enzyme Conjugate Buffer.	Dilute conjugate according to the Directions for Use.
Wash Buffer	Wash Buffer containing Sodium Azide has been used instead of the wash buffer provided in the kit.	The Wash Buffer in the kit is optimized for use with Mercodia ELISA kits. Washing solution from other companies may contain Sodium Azide, which can deactivate the enzyme. We therefore recommend using the Wash Buffer included in the kit.

Sources of error	Cause	Background/Suggestions
Calibrator, sample	No sample or Calibrator was added to the well.	It can, especially for assays with low sample volume, be difficult to see if Calibrator/Sample has been added to each well. Be careful not to miss a well when pipetting calibrators and samples.
Sodium Azide	Sample contains Sodium Azide.	Sodium Azide can deactivate the enzyme.

Sources of error	Cause	Background/Suggestions
Calibrator, sample	No sample or Calibrator was added to the well.	It can, especially for assays with low sample volume, be difficult to see if Calibrator/Sample has been added to each well. Be careful not to miss a well when pipetting calibrators and samples.
Sodium Azide	Sample contains Sodium Azide.	Sodium Azide can deactivate the enzyme.
Plate reader lamp	The intensity of the ELISA plate reader lamp is too low.	Check lamp efficiency and plate alignment. This can be done according to the Tech note 34-0107 Instruction for plate reader (Technical Library).
Absorbance	Absorbance was read with another wavelength than 450.	Check that the wavelength is set to 450 nm and that the right filter is in the plate reader.
Plate shaker	Plate shaker amplitude is too low.	Plate shakers should have a speed of approximately 700-900 cycles per minute (rpm). Low frequency results in poor calibration curve.
Pipettes	Pipettes are not calibrated, resulting in low sample/calibrator volume.	Pipettes should be calibrated and cleaned regularly.
Incubation time	Incubation time is too short.	Do not prolong or shorten incubations.
Preparation of enzyme conjugate solution	Over dilution of enzyme conjugate solution.	Dilute enzyme conjugate solution according to Directions for Use.
Incorrect wash	Soak was included in the wash procedure	Program the plate washer to aspirate immediately after the dispensing of Wash Buffer. If washing manually hold the plate vertically. When washing manually use a squirt bottle, see Tech note 34-0106 Instruction for manual washing (Technical Library).

Sources of error	Cause	Background/Suggestions
Calibrator, sample	No sample or Calibrator was added to the well.	It can, especially for assays with low sample volume, be difficult to see if Calibrator/Sample has been added to each well. Be careful not to miss a well when pipetting calibrators and samples.
Sodium Azide	Sample contains Sodium Azide.	Sodium Azide can deactivate the enzyme.
Washing	Insufficient washing.	Plate washer should be checked and served periodically. When washing manually use a squirt bottle, see Tech note 34-0106 Instruction for manual washing (Technical Library).
Contamination of Calibrator 0, Assay Buffer or Conjugate Buffer	Contamination of Calibrator 0, Assay buffer or Enzyme Conjugate Buffer with Calibrators, Controls or samples.	Always use clean pipette tips, beakers etc. in order to avoid contamination.
Contamination of substrate	Substrate might be contaminated with enzyme conjugate.	Containers and pipette tips used for enzyme conjugate solution should be kept separate from the substrate.
Substrate TMB	Substrate TMB has been exposed to light.	Substrate TMB is light sensitive. Uncovered Substrate can cause increased background if left out in the light for only a couple of minutes.
Dried solution on the bottom of the plate	Dried wash solution under the plate, on the outside of the wells.	Dampen a tissue with alcohol and carefully wipe off the bottom of the plate. Re-measure immediately.

Sources of error	Cause	Background/Suggestions
Calibrator, sample	No sample or Calibrator was added to the well.	It can, especially for assays with low sample volume, be difficult to see if Calibrator/Sample has been added to each well. Be careful not to miss a well when pipetting calibrators and samples.
Sodium Azide	Sample contains Sodium Azide.	Sodium Azide can deactivate the enzyme.
Washing	Insufficient washing.	Plate washer should be checked and served periodically. When washing manually use a squirt bottle, see Tech note 34-0106 Instruction for manual washing (Technical Library).
Pipettes	Pipettes are contaminated or out of function.	Pipettes should be calibrated and cleaned regularly.
Dried solution on the bottom of the plate	Dried wash solution under the plate, on the outside of the wells.	Dampen a tissue with alcohol and carefully wipe off the bottom of the plate. Re-measure immediately.
Bubbles in the wells when the plate was read	Pipetting error	Use repeating or multi-channel pipettes when adding substrate and Stop Solution. Use reverse pipetting to avoid bubbles.
Greenish colour in the wells	Insufficient stop of Substrate TMB reaction	Mix the plate immediately after adding Stop Solution to the wells.
Plate shaker	Plate shaker amplitude is too low.	Plate shakers should have a speed of approximately 700-900 cycles per minute (rpm) orbital movement. Low frequency results in poor calibration curve.

Sources of error	Cause	Background/Suggestions
Calibrator, sample	No sample or Calibrator was added to the well.	It can, especially for assays with low sample volume, be difficult to see if Calibrator/Sample has been added to each well. Be careful not to miss a well when pipetting calibrators and samples.
Sodium Azide	Sample contains Sodium Azide.	Sodium Azide can deactivate the enzyme.
Reconstitution volume (for kits with lyophilized calibrators or controls)	The lyophilized calibrators have been reconstituted with a smaller volume than stated in the Directions for Use.	Reconstitute with the correct volume of redistilled water as stated in the Direction for Use. Mix well before using. Check that there is no un-dissolved material left on the stopper of the bottle.
Hook effect	The sample contains a very high amount of antigen and is therefore given a falsely low result because of the hook effect.	Dilute and re-run the samples.
Dilution	The samples and/or controls have been diluted too much.	Always use the volumes stated in the Direction of Use for samples, calibrators, controls and enzyme conjugate solution. Check that pipettes are calibrated.

Sources of error	Cause	Background/Suggestions
Calibrator, sample	No sample or Calibrator was added to the well.	It can, especially for assays with low sample volume, be difficult to see if Calibrator/Sample has been added to each well. Be careful not to miss a well when pipetting calibrators and samples.
Sodium Azide	Sample contains Sodium Azide.	Sodium Azide can deactivate the enzyme.
Reconstitution volume (for kits with lyophilized calibrators or controls)	The lyophilized calibrators have been reconstituted with a larger volume than stated in the Directions for Use.	Reconstitute with the correct volume of redistilled water as stated in the Direction for Use. Mix well before using.
Dilution	The samples and/or controls have not been diluted enough.	Always use the volumes recommended for samples, Calibrators, Controls and enzyme conjugate solution. Check that pipettes are calibrated.

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