Olink Target 48 High Multiplex Immunoassay Panels Instructions

Olink Target 48 High Multiplex Immunoassay Panels

Incubation

1. Prepare the Incubation mix in a microcentrifuge tube according to the table below.

1. Vortex and spin down the Incubation mix. Transfer 23 µL of the Incubation mix to each well of a new 8-well strip.
2. Transfer 3 μL of Incubation mix to each well of the first 6 columns of a 96-well plate by reverse pipetting and name the plate Incubation Plate.
3. Add 1 µL of each sample using a multi-channel pipette to the bottom of the well, 1 µl of Sample Control to the three top wells (yellow), 1 µL of Negative Control to two wells (red), and 1 µL of Calibrators to three wells (green), according to the plate layout.
4. Seal the plate with an adhesive plastic film, spin at 400 – 1000 x g, 1 min at room temperature. Incubate overnight at +4 °C.

Extension

1. Prepare an extension mix according to the table below.
2. Bring the Incubation Plate to room temperature, spin at 400 – 1000 x g for 1 min. Preheat the PCR machine.
3. Vortex the Extension mix and pour it into a multichannel pipette reservoir.
4. Start a timer for 5 min and transfer 96 µL of Extension mix to the upper parts of the well walls of the Incubation Plate by using reverse pipetting.
5. Seal the plate with a new adhesive plastic film, use the MixMate® to vortex the plate at 2500 rpm for 30 sec, ensuring that all wells are mixed, and spin down.
6. Place the Incubation Plate in the thermal cycler and start the PEA program. (50 °C 20 min, 95 °C 5 min (95 °C 30 sec, 54 °C 1 min, 60 °C 1 min) x 17, 10 °C hold)

Detection

1. Prepare and prime an Olink® 48.48 IFC for Protein Expression. Briely, inject one control line fluid syringe into each accumulator on the chip, and then prime the IFC on the Olink® Signature Q100 following the instructions on the instrument screen.
2. Thaw the Primer Plate, vortex and spin briefly.
3. Prepare a Detection mix according to the table below.
4. Vortex the Detection mix and spin briefly and add 46 µL of the mix to each well of an 8-well strip.
5. Transfer 7.2 μL of the Detection mix to each well of column 1-6 in a new 96-well plate by reverse pipetting, and name it Sample Plate.
6. Remove the Incubation Plate from the thermal cycler, spin down the content and transfer 2.8 µL to the Sample Plate, using forward pipetting.
7. Seal the plate with an adhesive film, vortex and spin both at 400 – 1000 x g, 1 min at room temperature.
8. Transfer 5 μL from each well of the Prime Plate and 5 μL of the Sample Plate into the primed 48.48 IFC left and right inlets, respectively. Use reverse pipetting and change tips after each primer or sample. Do not leave any inlets empty.
9. Remove bubbles and load the chip in the Olink Signature Q100 and follow the instructions on the instrument screen.
10. Run the IFC on the Olink Signature Q100.
11. Carefully remove the adhesive film from the Primer Plate to avoid contamination between wells.
12. Transfer 5 µL of each primer using reverse pipetting from each well in position 1 A-H (green) to the inlets in the first column on the left side of the IFC (green). Change pipette tips after each column. When using an eight-channel pipette every other inlet will be filled according to the image.

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• For research use only. Not for use in diagnostic procedures.
  

• This product includes a license for non-commercial use. Commercial users may require additional licenses. Please contact Olink Proteomics AB for details.

• There are no warranties, expressed or implied, which extend beyond this description. Olink Proteomics AB is not liable for property damage, personal injury, or economic loss caused by this product.
  

• Olink® is a registered trademark of Olink Proteomics AB.
  

• © 2018–2022 Olink Proteomics AB. All third-party trademarks are the property of their respective owners.
  

• Olink Proteomics, Dag Hammarskjölds väg 52B , SE-752 37 Uppsala, Sweden
  

• 1126, v1.1, 2022-05-05

Documents / Resources

| Olink Target 48 High Multiplex Immunoassay Panels [pdf] Instructions
Target 48, High Multiplex Immunoassay Panels, Target 48 High Multiplex Immunoassay Panels, Immunoassay Panels, Panels, Target 48 Immunoassay Panels, Target 48 Panels
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| Olink Target 48 High Multiplex Immunoassay Panels [pdf] Instructions
Target 48, High Multiplex Immunoassay Panels, Target 48 High Multiplex Immunoassay Panels, Immunoassay Panels, Panels, Target 48 Immunoassay Panels, Target 48 Panels

References

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