Mouse/Rat beta-Amyloid (1-42) ELISA Assay Kit (plasma)

$1,300.00

The Eagle Biosciences Mouse/Rat beta-Amyloid (1-42) ELISA Assay Kit is intended  for the quantification of mouse/rat beta-Amyloid (1 – 42) in serum and plasma. The Eagle Biosciences Mouse/Rat beta-Amyloid (1-42) ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80981 Categories: , ,

Mouse/Rat beta-Amyloid (1-42) ELISA Assay Kit (plasma)

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.05 pg/ml
Dynamic Range: 1.56-100 pg/mL
Incubation Time: 3 hours
Sample Type: Serum, Plasma
Sample Size: 100 µL

Additional Information

Assay Background

Alzheimer’s Disease (AD) is the most common neurodegenerative disorder in elderly people. It has been demonstrated that AD has biological causes and is characterized by the presence of senile plaques and neurofibrillary tangles mainly in cerebral cortex and hippocampus brain regions. Beta-Amyloid (1-40) (Aβ40) and beta-Amyloid (1-42) (Aβ42) are the main components of the above plaques; however, other forms of beta-Amyloid peptides are also present. Both peptides are cleaved from the Amyloid Precursor Protein (APP) by β-secretase and γ-secretase enzymes. Many studies suggest that Aβ42 or/and Aβ43 are required to initiate formation of amyloid plaques and neurofibrills that leads to the neurodegeneration, while Aβ40 is less neurotoxic.

Assay Principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for mouse/rat Aβ (38-42) has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells. After washing away any unbound substances, the HRP-labeled mouse/rat Aβ (1-16) antibody is added to each well and incubate. Following a washing to remove unbound substances, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Aβ 1-42 bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm. The concentration of Aβ 1-42 in the sample is then determined by comparing the O.D of samples to the standard curve. The concentration of antigen is directly proportional to the optical density measured in the wells.

Assay Procedure

1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.

2. Add 100 μl of standards (S0-S8) and samples in duplicates into wells. Add 100 μl Assay Buffer in the other wells as reagent blank.

3. Cover the plate and incubate the plate at 4°C for overnight.

4. Aspirate each well and wash, repeating the process 6 times for a total 7 washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Keep the wash buffer in the wells for 15-30 seconds before remove it. Complete removal of liquid at each wash step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.

5. Add 100 μl of 1X HRP-labeled detection antibody into each well (except reagent blank), and incubate the plate at 4°C for 60 min in dark.

6. Aspirate each well and wash, repeating the process 5 times for a total 6 washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Keep the wash buffer in the wells for 15-30 seconds before remove it. Complete removal of liquid at each wash step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.

7. Add 100 μl of TMB Reagent to each well. Incubate for 10-15 minutes at room temperature in dark.

8. Add 50 μl of Stop Solution to each well. The color of the solution should change from blue to yellow.

9. Read the OD with a microplate reader at 450nm immediately.

Manual

Product Manual