Human SPARC 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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100 µL Assay Diluent

3.   Add 100 µL of Assay Diluent to each well.

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100 µL Standard, Control, or Sample

4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
5.   Aspirate each well and wash, repeating the process 5 times for a total of 6 washes.

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200 µL cold Conjugate

6.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour on the benchtop.
7.   Aspirate and wash 6 times.

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200 µL Substrate Solution

8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

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50 µL Stop Solution

9.   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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