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MedFrontier FGF23 (Human Intact FGF23) ELISA Assay Kit

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$1,050.00
SKU:
63278

 Product Description

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MedFrontier Human Intact FGF23 ELISA Assay Kit:
For Research Use Only
Size:  1x96 wells
Dynamic Range:  15 - 3000 pg/mL 
Incubation Time:  2.5 Hours
Sample Type:  Serum
Sample Size: 20 μL

Intended Use

The MedFrontier Human Intact FGF23 ELISA Assay Kit is a sandwich ELISA that measures ONLY the full-length active form (intact form). FGF23 is being researched in the context of X-linked hypophosphatemic rickets, mineral bone disorder (MBD), chronic kidney disease (CKD), tumor-induced osteomalacia and hyperphosphatemia. MedFrontier FGF23 is for Research Use Only and is not intended for use in diagnostic or therapeutic procedures.

Assay Background

FGF23 (Fibroblast Growth Factor 23) is a protein belonging to the fibroblast growth factor family. FGF23 is involved in the regulation of phosphorus metabolism. FGF23 has a molecular weight of approximately 32 kDa. FGF23 is produced in bone cells. In vivo, FGF23 is secreted into circulation. A full-length active FGF23 protein may undergo proteolytic cleavage to generate an inactive c-terminal fragment. This mechanism is thought to play a key role in controlling the concentration of circulating bioactive FGF23 in vivo.

Assay Principle

This sandwich ELISA kit uses two anti-human FGF23 mouse monoclonal antibodies. When serum is added to a plate well coated with anti-human FGF23 mouse monoclonal antibodies, FGF23 is captured by the immobilized antibodies (1st reaction). After the 1st reaction, the plate is washed. Then, ALP-labeled anti-human FGF23 mouse monoclonal 2nd antibodies against FGF23 react to FGF23 antigens captured by the immobilized antibodies (2nd reaction). After the 2nd reaction, the plate is washed and light detection is performed after adding the luminescence reagent. Each active well of the plate is measured using a luminescence microplate reader and relative light units (RLU) are obtained. The concentration of FGF23 in serum is calculated with a standard curve generated using the FGF23 Standard 1 to 6.

Assay Procedure

  1. Add 80 μL of Sample Dilution solution to each well of the Monoclonal Antibody (MAb)-Coated MicrotiterPlate and 20 μL each of FGF23 Standard 1 to 6, FGF23 Control L & H, or serum samples.
  2. Seal the plate and set it aside for 90 minutes at room temperature (18–25°C).
  3. After 90 minutes, remove the plate seal, aspirate the reaction solution, and wash each well 5 timeswith 500 μL of Wash Buffer. It is recommended to overflow the wells with wash buffer.
  4. Add 100 μL of ALP-Labeled Monoclonal Antibody (MAb) Reagent to each well. Seal the plate and set itaside for 90 minutes at room temperature (18–25°C).
  5. After 90 minutes, remove the plate seal, aspirate the reaction solution, and wash each well 5 timeswith 500 μL of Wash Buffer. It is recommended to overflow the wells with wash buffer.
  6. Add 100 μL of ALP Substrate (LumigenTM APS-5) solution to each well, and block out light for 1 minuteat room temperature (18–25°C).
  7. Measure relative light units (RLU) within 10 minutes.

Typical Standard Curve

standard-curve.png

References

  • Yuji Yamazaki, et al. (2002) J Clin Endocrinol Metab 87 (11):4957–4960
  • Itsuro Endo, et al. (2008) Bone 42:1235–1239
  • Vincent S. Tagliabracci, et al. (2014) Proc Natl Acad Sci USA 111(15):5520–5525.
  • Chikako Nakano, et al. (2012) Clin J Am Soc Nephrol 7 
  • Hugo Diniz, et al. (2013) Nefrologia 33:835–844.
  • Mariano Rodriguez, et al. (2012) Nefrologia 32 (3):275–278.
  • Myles Wolf, et al. (2012) Kidney Int 82 (7):737–747.
  • In-house data from Kyowa Medex Co., Ltd.

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Eagle Biosciences, Inc.
20A NW Blvd., Suite 112
Nashua, NH 03063
Email: info@eaglebio.com
Toll Free: 866-411-8023
International: +617-419-2019

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