The IMMUNO-TEK ELISA Construction System is an easy to use kit that enables scientists to rapidly prepare high quality ELISA assays. ELISA assays prepared with the IMMUNO-TEK ELISA Construction System are of comparable quality to those manufactured by the world’s leading immunodiagnostic companies. The kit contains specially screened microtiter plates and all reagents necessary to coat the plates with antigen or antibody. Microtiter plates prepared with the IMMUNO-TEK ELISA Construction System have a low background, high sensitivity and may be stored desiccated for one to two years with no loss of activity.

For convenience, all components of the IMMUNO-TEK ELISA Construction System may be ordered individually from ZeptoMetrix. This is to meet the needs of scientists who wish to further optimize their ELISA assays through the use of special microtiter plates or coating conditions.

Materials Supplied

• ZeptoBind, 60 ml

• ZeptoBlock, 100 ml

• ZeptoCoat, 60 ml

• Microtiter Plates, 5 each

• Sealable Bags, 5 each

• Desiccant Pillows, 5 each

• Plate Sealers, 5 each

Materials Required but Not Supplied

• Disposable gloves.

• Validated adjustable micropipets, single and multichannel.

• Test tubes and racks for preparing specimen and control dilutions.

• Graduated cylinders and assorted beakers.

• Validated automatic microplate washer or manual vacuum aspiration equipment.

• Validated incubator for 37°C ±1°C.

• Validated microplate reader.

• Timer.

• 1% sodium hypochlorite as disinfectant. May be prepared from household bleach.

• Distilled or deionized water.

Allow all reagents to reach room temperature before use. 

Step 1: Dissolve antigen or antibody in ZeptoBind. Prepare enough to add 100 µl to each well to be coated.

Step 2: Pipet 100 µl of antigen or antibody into each microtiter well.

Step 3: Cover plate with a plate sealer and incubate plate overnight at 4º C.

Step 4: Aspirate antigen or antibody.

Step 5: Pipet 200 µl of ZeptoBlock into each microtiter well.

Step 6: Cover microplate with a plate sealer and incubate for 2 hours at room temperature.

Step 7: Aspirate ZeptoBlock.

Step 8: Pipet 100 µl of ZeptoCoat into each microtiter well.

Step 9: Cover with a plate sealer and incubate 1 hour at room temperature.

Step 10: Aspirate ZeptoCoat and allow plate to dry for 4 hours at room temperature.

Step 11: Place plate and one desiccant pillow into a resealable bag and store plate until use. Plates prepared by this method will be stable for 12 to 24 months.

When possible, we recommend titrating the amount of antigen or antibody to be used in plate coating. We also recommend titrating the amout of antigen or antibody when using new lots of materials as there may be significant lot-to-lot variations in antigen or antibody preparations.

In such titrations, increasing amounts of antigens or antibodies will increase the signal, eventually reaching a plateau. Ideally, for an optimal assay one wishes to be at this plateau. Concentrations of antigen or antibody above that needed to achieve a plateau are not only wasteful but in some assays can lead to unacceptable backgrounds in the assay. For example, antigen concentrations from 1 to 10 µg/ml may yield steadily increasing signal but concentrations above 10 µg/ml show no further signal increase. In this case, 10 µg/ml would represent the plateau and the optimal coating concentration would be 10 µg/ml.

 DISSOLVE ANTIGEN OR ANTIBODY IN ZEPTOBIND

ADD 100 µl TO EACH WELL

INCUBATE OVERNIGHT AT 4º   C

ADD 200 µl ZEPTOBLOCK TO EACH WELL

ADD 100 µl ZEPTOCOAT TO EACH WELL

STORE AT 4º C