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Procedure:
Patients and Methods
1 - Dilute plasma samples with sample Buffer prior to adding to the
microtest wells. Serial dilutions were prepared as follows: 20 plasma to
1ml was added of sample buffer and mix; then add 20 ul ofthis initial
mixture to 1 ml ofsample buffer and mix; final dilution 1 : 2601.
2 - Add 50 ul ofPET Buffer to each microtest well.
3 - Add 20 ofeach Lp(a) standard (0 mg/I and 600 mg/l) to two separate
wells in duplicate. (standards was added without further dilution with
sample buffer). Add 20 ul ofeach diluted sample into a separate well. the
positions place the strips in the frame cover with a lid or plastic film.
Incubate for 60 minutes at room temperature on a micro-test plate shaker,
speed 500 pm
4 - Add 50 ul ofconjugate to each well using the 8-channel pipette .Cover
with lid or plastic film and incubate for 15 minutes at 25 CO with agitation
on a micro-test plate shaker.
5 - Empty the plate contents and wash the strips four times with PET
buffer: by filling the wells with PET buffer using a squeeze bottle, wait
for 1 minute, empty the plate and remove droplet by tapping the strips
and frame 4-5 times, face down, against absorbent material repeat do not
allow plate to dry out.
6 - Add 100 ul of substrate to each well using an 8-channel pipette or.
Start the stop watch. Incubate the strips with cover on the micro-test plate
shaker for 15 minutes at 25 C.
7 - Add 50 ul sulfuric acid to each well. Add this with the same speed and
order as you added the substrate . Agitate the plate for 5 minutes on a
micro-test plate shaker to allow complete mixing and stabilization of
color. The colored products is stable for 2 hours.
8 - Read the absorbance at 492 nm in a micro-test plate reader.
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