Bender MedSystems Instant ELISA® FAQs
General FAQs:
How many tests can I perform with an Bender MedSystems Instant ELISA®?
With an Instant ELISA® 128 tests can be performed. The number of tests results from the following composition:
- One 96 well plate designed only for samples
- 2x2 standard rows (= 2x16 wells) are separately packed per kit. Thus the kit can be used at two different time points.
It is possible to order more standard curves separately. The standard curves are lot depending. Please simply ask for availability before ordering.
Do I have to add reference material (standard) to perform a quantitative analysis?
No, the standard curve is already on the plate.
Do I have to add the second antibody for detection the test?
No, it's already on the plate.
Do I have to dilute my samples before testing?
Generally speaking, no, simply add the volume of sample as indicated. For some ELISAs an external pre-dilution will be required.
Is the reconstitution volume of the standards the same for different lot numbers?
For some Bender MedSystems Instant ELISAs® the reconstitution volume of the standard and blank wells may differ (these components are lot specific). Please do not use the manual on the website (general lot-unspecific information), but the manual in the Kit.
How do I have to calculate my samples?
The calculation factor is always stated in the manual.
An example for calculation of an Bender MedSystems Instant ELISA® with the dilution factor 1:2 is given here:
Prior to lyophilization the wells are filled with liquids of the respective components (standard, conjugate antibodies, diluents) in the correct volume and concentration. This volume of the components has to be reconstituted when the kit is assayed.
Calculation is based on the relation of standard and sample volume.
Sample wells: 150ul endvolume composed of 50ul sample, 50 ul diluent; 50 ul biotin conjugate. You have to add 100ul water to reconstitute the conjugate and diluent plus 50ul sample.
Standard wells: The standard wells for this kit are composed of diluted standard protein and 50ul biotin conjugate. Therefore you can omit the 50ul biotin conjugate volume for the calculation of the sample dilution. (The biotin conjugate contributes with equal volumes to standard and sample wells).
Correct calculation is 50ul sample diluted to 100ul volume therefore 1:2
The same is true for assays which have been diluted with other factors.
Troubleshooting (special for Bender MedSystems Instant ELISAs®)
How can I avoid high CV values?
- Use plate immediately after removal from -20°C. Do not put the plate outside of the foil and wait for a longer time before reconstitution of the samples. Otherwise the lyophilisate will absorb water depending on the humidity.
- Do not wait until pellets have completely dissolved before applying samples - the binding reaction in the standard strips starts immediately after addition of water!
- Do not try to dissolve pellets by pipetting up and down in the wells - some parts of the pellet could stick to the tip creating high CVs.
- Perform the washing step with at least 400µl of washing buffer as stated in the manual or fill the wells completely - otherwise any lyo-pellet residues sticking to the rim of the well will not be removed and create high CVs. Leave the wash buffer in the wells for a few seconds.
- Remove covers of the standard strips carefully so that all of the lyo-pellets remain in the wells.
