Apolipoprotein A1 is a protein that in humans is encoded by the APOA1 gene. This gene encodes apolipoprotein A-I, which is the major protein component of high density lipoprotein (HDL) in plasma. The encoded preproprotein is proteolytically processed to generate the mature protein, which promotes cholesterol efflux from tissues to the liver for excretion, and is a cofactor for lecithin cholesterolacyltransferase (LCAT), an enzyme responsible for the formation of most plasma cholesteryl esters. This gene is closely linked with two other apolipoprotein genes on chromosome 11. Defects in this gene are associated with HDL deficiencies, including Tangier disease, and with systemic non-neuropathic amyloidosis. Alternative splicing results in multiple transcript variants, at least one of which encodes a preproprotein. Sample Type: Cell culture supernates, serum, urine, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human APOA1/Apoa I. Sensitivity: <50pg/ml Range: 3.12ng/ml-200ng/ml Specificity: Recognizes natural and recombinant Human APOA1/Apoa I. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human APOA1/Apoa I BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for APOA1/Apoa I has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: D25-Q267) and test samples are added to the wells, a biotinylated detection specific for APOA1/Apoa I 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 APOA1/Apoa I amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human APOA1 antibody, 1x plate Lyophilized recombinant Human APOA1 standard, 2x200ng/tube Biotinylated anti-Human APOA1 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.
