Assay Procedure - peviva M30 Apoptosense ELISA Gebrauchsanweisung

Instructions for Use

Assay Procedure

The M30 Apoptosense ELISA should be performed at room temperature
(24 ± 3 °C).
1. Allow all reagents to reach room temperature before performing the assay.
Vortex all reagents prior to use.
2. Dissolve the Wash Tablet in fresh deionised water (see "Component Prepara-
tion").
3. Dilute the M30 Conjugate with M30 Conjugate Dilution Buffer (see "Compo-
nent Preparation") and mix.
4. Pipette 25 µL of M30 Standard (A – G), M30 Control Low, M30 Control High or
sample per well (duplicates are recommended).
5. Add 75 µL of the diluted M30 Conjugate solution to each well.
Note! Steps 4 and 5 should be performed sequentially without interruption
within 20 minutes.
6. Cover the wells with sealing tape or a microtiter plate lid.
7. Incubate on shaker for four (4) hours. Speed setting: 600 rpm.
8. Wash the plate in a plate washer five (5) times with 400 – 500 µL/well of
Wash Tablet solution (overflow wash)
or
Wash the plate manually, discarding the incubation solution and washing
the wells five (5) times with 250 µL of Wash Tablet solution. Avoid contami-
nation between wells.
9. Add 200 µL of TMB Substrate to each well. Incubate in darkness at room
temperature for 20 ± 1 minutes.
10. Add 50 µL of Stop Solution to each well. To ensure complete mixing of the
TMB Substrate and the Stop Solution, shake the microplate for 5 – 10 sec-
onds. Leave the microplate for 5 minutes before reading the absorbance.
11. Determine the absorbance at 450 nm in a microplate reader within 30 min-
utes and record the results.
12. Calculate the results as described in section "Calculation of Analytical Re-
sults".
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