Aliquot required volume of reagents and prepare before using.
Prepared reagents must be used immediately and avoid storing.

[ Preparation of Reaction Buffer ]

Make a 5-fold dilution of the Concentrated Reaction Buffer (G) in distilled water and prepare the required amount.
This Reaction Buffer is used for dilution of KS standards and samples, and is also used as a Blank Solution.

[ Preparation of HRP-Conjugated Antibody Solution ]

1)	Add 200 µL of HRP-Conjugated Antibody Diluent (F) to the contents of one vial of Concentrated HRP-Conjugated Antibody (C), and ensure it is fully dissolved (HRP-Conjugated Antibody Concentrate Solution).
2)	Dilute HRP-Conjugated Antibody Concentrate Solution with to make a 101-fold dilution in Reaction Buffer and mix well (dilute at the date of use). (Example preparation volume: 6 mL Reaction Buffer + 60 µL HRP- Conjugated Antibody Concentrate Solution)

[ Preparation of Washing Solution ]

Make a 5-fold dilution of the Concentrated Wash Solution (H) in distilled water to prepare the required amount.
Use the prepared Washing Solution for washing the microplate wells.

[ Preparation of Keratan Sulfate Standard Solutions ]

Use the prepared Reaction Buffer to dilute the Keratan Sulfate Standard (8 ng/mL) (B) and prepare five concentrations in a 1/2 dilution series (4, 2, 1, 0.5, and 0.25 ng/mL).
Use the Reaction Buffer as blank KS Solution (0 ng/mL).

[ Other reagents ]

Other kit components are ready to use.