Please find the link to the following protocol:
2-MICROGLOBULIN ENZYME IMMUNOASSAY(EIA) PROCEDURE
PRINCIPLE AND PURPOSE OF TEST:
2-Microglobulin (2-M) Enzyme immunoassay (EIA) can be used for the quantitative determination of 2-M in human serum and urine. ( References) 2-M EIA kit is available from ORGenTec. It is a solid phase enzyme immunoassay. The
microplate is coated with highly purified anti -human 2-M antibodies. Calibrators , prediluted patient samples , and controls are pipetted into the wells of microplate . After 30 minutes incubation the plate is washed with wash buffer solution to remove non-reactive components , after final wash
an anti-human 2-M-Horseradish peroxidase conjugate solution is added into all wells to recognize 2-M bound to the immobilized antibodies by forming the sandwich complexes. After 15 minutes incubation the plate is washed to remove unbound enzyme conjugate. A choromogenic solution
containing TMB is added into the wells. After 15 minutes incubation the color development is stopped by adding 1 M hydrochloric acid as stop solution. The absorbance of each well is read at 450 nm and standard curve is constructed to quantify 2-M concentration in the samples. The amount of color is directly proportional to the concentration of 2-M present in the sample.