• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Range

  156.00 - 10,000 pg/mL

• Sufficient Materials

  For five 96-well plates*

• Specificity

  Please see the

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Nephrin. The suggested diluent is suitable for the analysis of most cell culture supernate, cell lysate, serum, and plasma samples. Diluents for complex matrices, such as cell lysate, serum, and plasma, should be evaluated prior to use in this DuoSet.

Product Features

• Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time

• Development protocols are provided to guide further assay optimization

• Assay can be customized to your specific needs

• Economical alternative to complete kits

Kit Content

• Capture Antibody

• Detection Antibody

• Recombinant Standard

• Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # ) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # ), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # ), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # )

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

Preparation and Storage

• Storage

  Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Nephrin

Nephrin, also known as renal glomerulus-specific cell adhesion receptor and nephrosis 1 (NPHS1) is a 185 - 200 kDa type I transmembrane protein belonging to the immunoglobulin (Ig) superfamily. It is expressed on podocytes and is an essential component of the interpodocyte-spanning slit diaphragm complex. Nephrin forms cis-hetero-oligomeric complexes with Neph1, followed by trans-homophilic interaction with Nephrin on opposing cells. Mutations in the Nephrin gene is the pathogenic cause of congenital nephrotic syndrome. Mature human Nephrin contains a 1033 aa extracellular region and a 165 aa cytoplasmic tail. One potential soluble splice form of nephrin is known where aas 1056 - 1095 are deleted, eliminating the transmembrane region. Over aa 23 - 1029, human Nephrin shares 84% and 89% aa identity with mouse and canine Nephrin, respectively.

• Entrez Gene IDs:

  4868 (Human); 54631 (Mouse); 64563 (Rat);

• Aliases:

  CNF; Nephrin; nephrosis 1, congenital, Finnish type (nephrin); NPHNCNF; NPHS1; Renal glomerulus-specific cell adhesion receptor