Rat AGP ELISA Assay
The Rat AGP ELISA Assay is For Research Use Only
Size: 1×96 wells
Sensitivity: 3.245 ng/ml
Standard Range: 12.5 – 400 ng/ml
Incubation Time: 80 minutes
Sample Type: Rat Biological Fluids (serum and plasma)
Alternative Names: Alpha 1-Acid Glycoprotein, a1a glycoprotein
Sample Size: 100 µl
The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Alpha 1-Acid Glycoprotein present in samples reacts with the anti-Alpha 1-Acid Glycoprotein antibodies which have been adsorbed to the surface of polystyrene microtitre wells. After the removal of unbound proteins by washing, anti-AGP antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound AGP. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of AGP in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of AGP in the test sample. The quantity of AGP in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.
SPECIMEN COLLECTION AND HANDLING
Blood should be collected by venipuncture. The serum should be separated from the cells after clot formation by centrifugation. For plasma samples, blood should be collected into a container with an anticoagulant and then centrifuged. Care should be taken to minimize hemolysis, excessive hemolysis can impact your results. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze thaw cycles.
For any sample that might contain pathogens, care must be taken to prevent contact with open wounds.
2. Additives and Preservatives
No additives or preservatives are necessary to maintain the integrity of the specimen. Avoid azide contamination.
3. Known interfering substances
Azide and thimerosal at concentrations higher than 0.1% inhibits the enzyme reaction.