User Manual /96 tests - Simoco Diagnostics

Tianjin Bio-Enoche Engineering Co., Ltd. YNQ-SM-1206-1 A/1
Addition of substrate solution:
TMB solution: 100μL/well
37℃ for 15min
Stop the reaction:
Stopping solution: 50μL/well
Measurement:
Read OD at 450nm
9. SAMPLE TREATMENT
9.1. Add 300μL of serum into the centrifugal tube.
9.2. Add 100μL of sample treatment solution (R5) into the above centrifugal tube.
9.3. Vor tex the centrifugal tube for 10sec. Heat the tube at 100℃ for 3min in water bath.
9.4. Centrifuge the heated tube at 4℃ for 10min at 10,000×g.
9.5. Collect 60μL of supernatant for detection.
10. PROCEDURE
10.1. Bring all reagents to room temperature for 30min before test.
10.2. Take the microtiter strips out of the sealed bag (R1). Return the unused strips and seal the
bag tightly.
10.3. Prepare washing solution :
Dilute the concentrated washing solution (20×) at 1:19 ratio with DD H 2 O. The resultant
washing solution is stored at 2~8℃ for up to 2 weeks. Adequate washing solution should
be prepared for the entire test.
10.4. Prepare the polypropylene centrifugal tubes (0.6mL or 1.5mL) in advance.
10.5. Add 60μL of standards (a, b, c, d and e) and samples into each tube separately, and then
add 60μL of anti-galactomannan antibody.
10.6. Vortex all tubes well. incubate all the tubes at 37℃ for 30 min.
10.7. Pipette 100μL of above mixture into wells as below. Add 100μL of sample dilution into
one well as the substrate blank.
Well A
Substrate Blank
Well B
Standard a
Well C
Standard b
Well D
Standard c
Well E
Standard d
Well F
Standard e
Well G Sample 1…
10.8. Seal the microtiter plate with a plate sealer and incubate it at 37℃ for 30min.
10.9. Remove the plate sealer and shake out the incubation solution. Wash the wells 3 times
with 300μL/well washing solution each time. The soak time is 40sec. After the last wash,
invert the microtiter plate and dry it by tapping on the absorbent paper. Add 100μL of
conjugate into each well except the substrate blank.
10.10. Seal the microtiter plate with a plate sealer and incubate it at 37℃ for 30min.
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