Acetylcholine Receptor Autoantibody ELISA

Enzyme immunoassay for the quantitative determination of AChRAb in human serum.

INTENDED USE:The RSR Acetylcholine Receptor Autoantibody (AChRAb) ELISA kit is intended for use by professional persons only, for the quantitative determination of AChRAb in human serum. Autoantibodies to the acetylcholine receptor (AChR) are responsible for failure of the neuromuscular junction in myasthenia gravis. Measurement of these antibodies can be of considerable value in disease diagnosis and management.

GENERAL INFORMATION:RSR’s AChRAb ELISA depends on the ability of AChRAb in human serum to bind to similar sites on the AChR as various monoclonal antibodies such as MAb1 (coated on ELISA plate wells) and/or MAb2 and/or MAb3 (which are labelled with Biotin). In the absence of AChRAb a complex is formed between MAb1 coated on the plate wells, the AChR and MAb2- and MAb3-Biotin. MAb2- and MAb3-Biotin bound are then detected by addition of Streptavidin Peroxidase (SA-POD), which binds specifically to Biotin. Excess, unbound SA-POD is then washed away and addition of the peroxidase substrate 3,3’,5,5’ – tetramethylbenzidine (TMB) results in formation of a blue colour. This reaction is stopped by addition of stop solution causing the well contents to turn yellow. The absorbance of the yellow reaction mixture at 450 nm is then read using an ELISA plate reader. In the presence of AChRAb the formation of the MAb1-AChR-MAb2-/MAb3-Biotin complex is inhibited, resulting in less SA-POD being bound and a reduction in final absorbance at 450 nm. The higher the concentration of AChRAb in the test serum, the greater the inhibition of MAb-Biotin binding. When using the kit calibrators, the measuring interval is 0.45 – 20 nmol/L toxin bound.

KIT CHARACTERISTICS:
-Method: ELISA
- 96 tests
-Incubation Time / Conditions: 16-20h (2-8°C), 1 h (RT/shaker), 1 h (RT/shaker), 30 min (RT/shaker), 30 min (RT/dark)
- Standard Range: 0.5 - 20 nmol/l
- Sensitivity analytical: 0.25 nmol/l
- Final Sample: Volume 100 µl
- Sample Type: serum
- Isotope / Substrate: TMB 450 nm
- Internal Controls: 3
- Regulatory Status: CE