METHODS
1) Prepare a sufficient number of Antibody Coated Strip in a holder.
2) Add 50 μL of Assay Diluent into the well.
3) Add 50 μL of FGF-23 Standards or samples into the each well, and cover the plate with a plate sealer.
4) Incubate plate at room temperature for 2 hrs on a plate mixer.
5) Remove plate sealer. Aspirate the contents of each well. Wash each well 4 times with 300 μL of working Wash Buffer and completely aspirate the contents.
6) Add 100 μL of FGF-23 Conjugate into each well and cover the plate with a plate sealer.
7) Incubate plate at room temperature for 1 hr on a plate mixer.
8) Remove plate sealer. Aspirate the contents of each well. Wash each well 4 times with 300 μL of working Wash Buffer into each well and completely aspirate the contents.
9) Add 100 μL of Substrate into each well and cover the plate with a plate sealer and a aluminum foil.
10) Incubate plate at room temperature for 30 minutes.
11) Remove plate sealer and aluminum foil. Immediately add 100 μL of Stop Solution into each well. Shake the plate on the plate mixer for 1 minute.
12) Read the absorbance at 450 nm within 10 min by a microtiter plate reader.
     
 

Prepare all reagents and standards

   
  Add 50 μL of Assay Diluent to each well
   
  Add 50 μL of FGF-23 Standards or samples to each well
   
  IIncubate at room temperature for 2 hrs on a plate mixer
   
  Aspirate and wash 4 times
   
  Add 100 μL of FGF-23 Conjugate into each well
   
  Incubate at room temperature for 1 hr on a plate mixer
   
  Aspirate and wash 4 times
   
  Add 100 μL of Substrate into each well
   
  Incubate at room temperature for 30 minutes
Protect from light
   
  Add 100 μL of Stop Solution into each well
Read at 450 nm within 10 minutes
     
     

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