The β-Amyloid (1-42) BioAssay™ ELISA Kit is to be used for the in vitro quantitative determination of human b-amyloid 42 in samples (e.g., tissue culture medium, tissue homogenate, cerebrospinal fluid (CSF), etc.). The assay will recognize both natural and synthetic forms of human b-amyloid 42. Anti-human b-amyloid 42 antibody used in this kit is capable of selectively detecting b-amyloid 42 and not b-amyloid 40/ b-amyloid 43.
Test Principle:A2275-75R is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for the NH2-terminus of human b-amyloid has been coated onto the wells of the microtiter strips provided. Samples, including standards of known human b-amyloid content, control specimens and unknowns are pipetted into these wells, and co-incubated with a rabbit antibody specific for the COOH-terminus of the 1-42 b-amyloid sequence. This COOH-terminal sequence is created upon cleavage of the analyzed precursor. Bound rabbit antibody is detected by the use of a horseradish peroxidase-labeled anti-rabbit antibody. After removal of excess anti-rabbit antibody, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of human b-amyloid (1-42) present in the original specimen.
Kit Components:Microtiter plate, 1x96wells. Plate pre-coated with monoclonal antibody to N-terminus of beta-amyloidb-Amyloid 1-42 Standard, 1x1 vial. Lyophilized synthetic peptide. Refer to vial label for quantity and reconstitution volume
Standard Diluent Buffer, 1x60ml. Contains 15mM sodium azide and red dye
Detection Antibody, 1x6ml. Rabbit anti-human b-amyloid 42. Contains 15mM sodium azide and blue dye
IgG(HRP) (100X), 1x125ul. Anti-rabbit IgG conjugated to horseradish peroxidase. Contains 3.3mM thymol and yellow dye
HRP Diluent, 1x25ml. Contains 3.3mM thymol and yellow dye
Storage and Stability:Store kit components at 4°C. Stable for at least 6 months. Store reconstituted human b-amyloid 1-42 standard in aliquots at -70°C for up to 4 months, For maximum recovery of product, centrifuge the original vial prior to removing the cap.
