WAP four-disulfide core domain protein 2, also known as Human Epididymis Protein 4 (HE4), is a protein that in humans is encoded by the WFDC2 gene. This gene encodes a protein that is a member of the WFDC domain family. The WFDC domain, or WAP Signature motif, contains eight cysteines forming four disulfide bonds at the core of the protein, and functions as a protease inhibitor in many family members. This gene is expressed in pulmonary epithelial cells, and was also found to be expressed in some ovarian cancers. The encoded protein is a small secretory protein, which may be involved in sperm maturation.
Sample Type
Serum, urine, cell culture supernates, saliva, human milk, plasma (Heparin or EDTA).
Intended Use
Sandwich High Sensitivity BioAssay™ ELISA kit for Quantitative Detection of Human HE4.
Range
156pg/ml-10,000pg/ml
Specificity
Recognizes natural and recombinant Human HE4. There is no detectable cross-reactivity with other relevant proteins.
Test Principle
This Human HE4 BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for HE4 has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli; Immunogen sequence: E31-F124) and test samples are added to the wells, a biotinylated detection specific for HE4 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human HE4 amount of sample captured in plate.
Kit Components
96-well plate precoated with anti-Human WFDC2 antibody, 1x plate Lyophilized recombinant Human WFDC2 standard, 2x10ng/tube Biotinylated anti-Human WFDC2 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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