Quansys Q-Plex Chemiluminescent assay instructions
1. Before beginning the assay make sure the imager is setup and focused.
2. Also Reconstitute all reagents just before starting the assay and store at 4 degrees C until they are needed.
3. To make the antigen standard place 65ul of sample diluent into seven wells of a source plate or seven tubes. Place 65ul of the high dilution point of the antigen standard into the first well or tube with sample diluent to make a 1:2 dilution.
4. Mix the dilution point well then take 65ul from that well or tube and place it in the next well or tube and mix. Continue down all the dilution points to make the antigen standard dilution series with a 1:2 dilution.
5. Do not mix antigen standard into the 8th or last point of the curve as it will serve as the negative control with sample diluent only.
6. Dilute any samples in the sample diluent until the concentration of the proteins in the samples will fall within the range of the standard curve.
7. If possible do several different dilutions for each sample to make sure they will fall within the range of the standard curve.
8. Open the plate bag when you are ready to place the samples and antigen standards on the plate.
9. Add 30ul of sample or standard to each well of the assay plate. Make sure you load the samples and standard to the plate in a timely manner.
10. Cover the plate and place it on a plate shaker for one hour at room temperature.
11. Wash the plate with the wash buffer three times using an automatic plate washer or by hand.
12. If washing the plate by hand aggressively flick the solution out of the plate over a waste container before starting the wash protocol.
13. Using a multichannel pipette deposit 200-300 ul of wash buffer into each well used in the test.
14. Agressively flick the wash buffer out over a waste container. This washes the plate one time. Repeat this procedure two more times for a three time wash.
15. Add the detection to a tray or trough, then pipette 30ul into each well used in the test.
16. Cover the plate and place it on a plate shaker for one hour at room temperature.
17. Following the hour incubation, wash the plate three times. Bang out the plate after washing to remove residual buffer.
18. Add the 1X Streptavidin HRP to a tray our trough, then pipette 30ul into each well used in the test.
19. Cover the plate and place it on a plate shaker for 15 minutes at room temperature.
20. While the plate is incubating prepare the substrate by adding equal parts of substrate A and substrate B to a tray or trough and gently mix it well.
21. Wash the plate six times then bang the plate out to remove any residual buffer.
22. Add 40ul of the mixed substrate to each well used in the test being careful not to pipette bubbles into the well.
23. Image the plate immediately after substrate is added for optimal results. Wait no more then 15 minutes to commence imaging.
24. It is recommended that multiple images at different exposure lengths be taken to ensure the best results for each analyte.
1. Before beginning the assay make sure the imager is setup and focused.
2. Also Reconstitute all reagents just before starting the assay and store at 4 degrees C until they are needed.
3. To make the antigen standard place 65ul of sample diluent into seven wells of a source plate or seven tubes. Place 65ul of the high dilution point of the antigen standard into the first well or tube with sample diluent to make a 1:2 dilution.
4. Mix the dilution point well then take 65ul from that well or tube and place it in the next well or tube and mix. Continue down all the dilution points to make the antigen standard dilution series with a 1:2 dilution.
5. Do not mix antigen standard into the 8th or last point of the curve as it will serve as the negative control with sample diluent only.
6. Dilute any samples in the sample diluent until the concentration of the proteins in the samples will fall within the range of the standard curve.
7. If possible do several different dilutions for each sample to make sure they will fall within the range of the standard curve.
8. Open the plate bag when you are ready to place the samples and antigen standards on the plate.
9. Add 30ul of sample or standard to each well of the assay plate. Make sure you load the samples and standard to the plate in a timely manner.
10. Cover the plate and place it on a plate shaker for one hour at room temperature.
11. Wash the plate with the wash buffer three times using an automatic plate washer or by hand.
12. If washing the plate by hand aggressively flick the solution out of the plate over a waste container before starting the wash protocol.
13. Using a multichannel pipette deposit 200-300 ul of wash buffer into each well used in the test.
14. Agressively flick the wash buffer out over a waste container. This washes the plate one time. Repeat this procedure two more times for a three time wash.
15. Add the detection to a tray or trough, then pipette 30ul into each well used in the test.
16. Cover the plate and place it on a plate shaker for one hour at room temperature.
17. Following the hour incubation, wash the plate three times. Bang out the plate after washing to remove residual buffer.
18. Add the 1X Streptavidin HRP to a tray our trough, then pipette 30ul into each well used in the test.
19. Cover the plate and place it on a plate shaker for 15 minutes at room temperature.
20. While the plate is incubating prepare the substrate by adding equal parts of substrate A and substrate B to a tray or trough and gently mix it well.
21. Wash the plate six times then bang the plate out to remove any residual buffer.
22. Add 40ul of the mixed substrate to each well used in the test being careful not to pipette bubbles into the well.
23. Image the plate immediately after substrate is added for optimal results. Wait no more then 15 minutes to commence imaging.
24. It is recommended that multiple images at different exposure lengths be taken to ensure the best results for each analyte.