FGF-23 ELISA kit

Anti FGF-23 antibody - N-terminal　FGF-23　C-terminal - HRP labeled anti FGF-23 antibody

At the end of this reaction, the well is washed to remove unbound components.

In enzyme reaction, the sandwich complex immobilized on the well is incubated with a substrate solution and then measured by a spectrophotometric microtiter plate reader. The enzymatic activity of the complex bound to the well is directly proportional to the amount of FGF-23 in the sample.

A standard curve is generated by plotting the absorbance versus the each concentration of FGF-23 standard. The concentration of FGF-23 in the sample is determined from this curve.

	1)	FGF-23 Antibody coated Microplate（FGF-23 Microplate） 96 well microplate（12 strips of 8 well）coated with a murine monoclonal antibody against FGF-23.
	2)	HRP labeled FGF-23 Antibody（FGF-23 Conjugate） 12 mL of murine monoclonal antibody against FGF-23 conjugated to horseradish peroxidase, with preservatives. This reagent should be stored at 2-10℃ and protected from light.
	3)	FGF-23 Standards（Std1～Std7） 12 mL （Std1） and 0.5 mL（Std2～Std7） of buffered FGF-23 with preservative. 7 vials of FGF-23 in buffered base with preservative.
	4)	Assay Diluent（Assay Diluent） 12 mL of buffer.
	5)	Color Reagent（Substrate） 12 mL of tetramethylbenzidineTMBZ）with urea hydrogen peroxide.
	6)	Wash Buffer Concentrate（Wash Buffer (×5)） 90 mL of 5-fold concentrated solution of buffered surfactant with preservative. Before using, dilute the contents to 5-fold with deionized water and mix well.
	7)	Stop Solution（Stop Solution） 12 mL of 0.5 mol/L-Sulfuric acid.
	8)	Plate Sealer Three included in kit.
		＊These reagents should be stored at 2－10 ℃ in dark.
		Before using bring all reagents to room temperature.       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