The M30-Apoptosense®
assay should be performed at room temperature (20±4°C). Bring all
reagents and samples to room temperature before performing the assay.
- Define the number of Coated Microstrips
needed and identify the reaction wells according to the test protocol for
Standards, Controls and Samples.
- Preparation of the HRP Conjugate Solution:
Dilute a proper amount of the HRP Conjugate Concentrate with Conjugate
Dilution Buffer (see Table 1 for guidance).
Note! The diluted HRP Conjugate Solution should be used at the time of
preparation, but is stable for 2 weeks after preparation if stored at
2-8°C in the dark.
- Pipette 25 µl Standard, Control or Sample to their assigned wells.
Without interruption, add 75 µl of the HRP Conjugate Solution to each well of
the Coated Microstrip(s).
- Agitate on shaker for 4 hours. For optimal speed settings,
see section 7: Materials required but not supplied.
- Preparation of Wash Solution:
Dilute the 50 ml Wash Solution Concentrate in 450 ml of fresh distilled water.
- Remove the fluid from the wells and wash each well 3 times with
Wash Solution. Avoid contamination between wells.
- Add 200 µl of TMB Substrate Solution to each well and incubate in
darkness for 20±1 minutes.
- Stop the reaction by adding 50 µl of the Stop Solution. Corrosive!
- To ensure complete mixing of the TMB Substrate Solution and the Stop
Solution, agitate the microplate on a Microplate Shaker for 5-10 seconds.
- Wait 5 minutes before reading.
- Measure the absorbance at 450 nm in a Microplate Reader and record
the results.
