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Nephrin ELISA Kit (NPHS1)

Full Name: Nephrin ELISA Kit (NPHS1)
Reactivity: Human
Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
Sensitivity: 0.094 ng/ml

INTRODUCTION

Nephrin (NPHS1) is an integral membrane protein that plays a critical role in the kidney as a structural component of the slit diaphragm. The slit diaphragm forms specialized intercellular junctions connecting podocyte foot processes in the glomerulus, thus comprising a key part of the filtration barrier. Loss of nephrin leads to massive proteinuria and nephrotic syndrome. The NPHS1 gene encodes nephrin, and mutations in this gene were first discovered in congenital nephrotic syndrome of the Finnish type, an autosomal recessive kidney disorder presenting in infancy. Since then, NPHS1 mutations have been found in steroid-resistant nephrotic syndrome manifesting in childhood worldwide. The most common mutation is the Finnish founder mutation Fin-major, but many other nonsense, frameshift, and missense mutations have also been identified.

As a transmembrane protein of the immunoglobulin superfamily, nephrin’s extracellular domain contains IgG-like motifs and a fibrinogen-like motif, allowing nephrin molecules on neighboring podocytes to interact. Nephrin’s short intracellular domain connects to the podocyte cytoskeleton via adaptor proteins like Nck and podocin. This link is essential for maintaining the filtration barrier integrity.

INTENDED USE

Human nephrin ELISA kit can measure concentrations of NPHS1 (nephrin, NPHS1, CNF, renal glomerulus-specific cell adhesion receptor, NPHNCNF) present is plasma, serum, biological fluids or tissue homogenate samples.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • One 96-Well Plate: Pre-coated with anti-NPHS1 antibody.
  • Standards: Lyophilized recombinant.
  • Sample/Standard Dilution Buffer.
  • Biotinylated-labelled Antibody.
  • Antibody Dilution Buffer.
  • HRP-Streptavidin Conjugate (SABC).
  • SABC Dilution Buffer.
  • TMB Substrate.
  • Wash Buffer (25x).
  • Plate Sealer.
  • Product Instructions.

TYPICAL RESULTS

For this nephrin ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.156, 0.312, 0.625, 1.25, 2.5, 5, 10 ng/ml.
Reactivity: Human
Sensitivity: 0.094 ng/ml
Range: 0.156 – 10 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for NPHS1, no cross reactivity or interference between NPHS1 and analogues was detected.
– Recovery: Serum (89 – 99%), EDTA Plasma (91 – 97%), Heparin Plasma (90 – 105%).
– Linearity: Serum (90 – 103%), EDTA Plasma (84 – 94%), Heparin Plasma (81 – 96%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

  1. Nephrin – a biomarker of early glomerular injury. Biomark Res. (2014) 2: 21. Kandasamy Y., et al.
  2. Expanding the spectrum of NPHS1-associated disease. Kidney Int. (2009) 76 (12): 1221-3. Pollak M.R.
  3. Nephrin gene (NPHS1) in patients with minimal change nephrotic syndrome (MCNS). Kidney Int. (2004) 65 (5): 1856-63. Lahdenkari A.T., et al.
  4. Nephrin in human lymphoid tissues. Cell Mol Life Sci. (2006) 63 (4): 498-504. Aström E., et al.

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