High Sensitive Apolipoprotein A-I (apoA1) ELISA Assay Kit

$550.00

The Eagle Biosciences High Sensitive Apolipoprotein A-I (apoA1) ELISA Assay Kit is intended for the quantification of Human Apolipoprotein A1 in serum, plasma, cell culture supernatants. The Eagle Biosciences Apolipoprotein A-I (apoA1) ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80132 Categories: , ,

High Sensitive Apolipoprotein A-I (apoA1) ELISA Assay Kit

For Research Use Only

Size: 1 x 96 wells
Sensitivity: 0.28 pg/ml
Dynamic Range: 1.56-100 pg/ml
Incubation Time: 3 hours
Sample Type: Brain Extract, Serum, Plasma, Cell Culture Supernatants
Sample Size: 100 µl

Additional Information

Assay Background

Apolipoprotein A I promotes cholesterol efflux from tissues to the liver for excretion. Apolipoprotein A I is the major protein component of high density lipoprotein (HDL) in the plasma. Synthesized in the liver and small intestine, it consists of two identical chains of 77 amino acids; an 18 amino acid signal peptide is removed co-translationally and a 6 amino acid propeptide is cleaved post-translationally. Apolipoprotein A I is a cofactor for lecithin cholesterolacyltransferase (LCAT) which is responsible for the formation of most plasma cholesteryl esters. Defects in the Apolipoprotein A I gene are associated with HDL deficiency and Tangier disease.

Assay Principle

This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for apoA1 has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any apoA1 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-conjugated antibody specific for apoA1 is added to each well and incubate. Following a washing to remove unbound substances, streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of apoA1 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 ±2nm. The concentration of apoA1 in the sample is then determined by comparing the O.D of samples to the standard curve.

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, samples and zero controls (standard diluent buffer) into wells. Incubate for 1.5 h at 36 °C.
  3. Aspirate each well and wash, repeating the process four times for a total five washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  4. Add 100 μl 1X Antibody conjugate into each well. Cover wells and incubate for 1 hour at 36°C.
  5. Aspirate each well and wash as step 3.
  6. Add 100 μl of 1X HRP-Streptavidin solution to each well. Cover wells and incubate for 30 minutes at 36°C.
  7. Aspirate each well and wash as step 3.
  8. Add 100 μl of TMB Reagent to each well. Incubate for 15 minutes at 36°C in dark.
  9. Add 100 μl of Stop Solution to each well. The color of the solution should change from blue to yellow.
  10. Read the OD with a microplate reader at 450nm immediately.

Typical Standard Curve

Manual

Product Manual