Apolipoprotein E (APOE) is a class of apolipoprotein found in the chylomicron and Intermediate- density lipoprotein (IDLs) that is essential for the normal catabolism of triglyceride-rich lipoprotein constituents. In peripheral tissues, APOE is primarily produced by the liver and macrophages, and mediates cholesterol metabolism in an isoform- dependent manner. In the central nervous system, APOE is mainly produced by astrocytes, and transports cholesterol to neurons via APOE receptors, which are members of the low density lipoprotein receptor gene family. This protein is involved in Alzheimers disease and cardiovascular disease. Sample Type: Cell culture supernates, serum, urine, saliva, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human APOE/Apolipoprotein E. Sensitivity: <50pg/ml Range: 3.12ng/ml-200ng/ml Specificity: Recognizes natural and recombinant Human APOE/Apolipoprotein E. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human APOE/Apolipoprotein E BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for APOE/Apolipoprotein E has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: K19-H317) and test samples are added to the wells, a biotinylated detection specific for APOE/Apolipoprotein E 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 APOE/Apolipoprotein E amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human APOE antibody, 1x plate Lyophilized recombinant Human APOE standard, 2x200ng/tube Biotinylated anti-Human APOE 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 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
