Human LIF Quantikine ELISA Kit

Assay Procedure

1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

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50 µL Assay Diluent for Serum & Plasma Samples Only

3.   For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.

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200 µL Standard, Control, or Sample

4.   Add 200 µL of Standard, control, or sample to each well.
5.   For Serum & Plasma Samples: Cover with a plate sealer, and incubate at 37 °C for 2 hours.
   For Cell Culture Supernate Samples: Cover with a plate sealer, and incubate at room temperature for 2 hours.
6.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

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200 µL Conjugate

7.   Add 200 µL of Conjugate to each well.
8.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
   For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
9.   Aspirate and wash 3 times.

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200 µL Substrate Solution

10. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

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50 µL Stop Solution

11. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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