Mumps IgG ELISA Kit

Mumps virus

The Mumps IgGantibody test kit is based on the principle of the enzyme immunoassay (EIA).Mumps antigen is bound on the surface of the microtiter strips. Dilutedpatient serum or ready-to-use standards are pipetted into the wells of themicrotiter plate. A binding between the IgG antibodies of the serum and the immobilizedMumps antigen takes place. After a one hour incubation at room temperature, the plate is rinsed with diluted wash solution, in order to remove unboundmaterial. Then ready-to-use anti-human-IgG peroxidase conjugate is added and incubatedfor 30 minutes. After a further washing step, the substrate (TMB) solution ispipetted and incubated for 20 minutes, inducing the development of a blue dyein the wells. The color development is terminated by the addition of a stop solution, which changes the color from blue to yellow. The resulting dye is measuredspectrophotometrically at the wavelength of 450 nm. The concentration of theIgG antibodies is directly proportional to the intensity of the color.

1. 5 uL-, 100 uL-and 500 uL micro- and multichannel pipets 2. Microtiter PlateReader (450 nm) 3. Microtiter PlateWasher 4. Reagent tubes forthe serum dilution 5. Bidistilled water

I-ELISA

1. ReagentAnd Sample Preparation WashingSolution: dilute before use1+9 with distilled water. If during the cold storage crystals precipitate, the concentrate should be warmed up at 37C for 15 minutes. 1) Strictadherence to the protocol is advised for reliable performance. Any changes ormodifications are the responsibility of the user. 2) Allreagents and samples must be brought to room temperature before use, butshould not be left at this temperature longer than necessary. 3)Standards and samples should be assayed in duplicates. 4) Astandard curve should be established with each assay. 5) Returnthe unused microtiter strips to the plastic bag and store them dry at 2-8oC. 2.Assay Steps 1) Preparea sufficient amount of microtiter wells for the standards, controls andsamples in duplicate as well as for a substrate blank. 2) Pipet100 uL each ofthe diluted (1:101) samples and the ready-to-use standards and controlsrespectively into the wells. Leave one well empty for the substrate blank. 3) Coverplate with the enclosed foil and incubate at room temperature for 60 minutes. 4) Emptythe wells of the plate (dump or aspirate) and add 300 uL ofdiluted washing solution. This procedure is repeated totally three times.Rests of the washing buffer are afterwards removed by gentle tapping of themicrotiter plate on a tissue cloth. 5) Pipet100 uL each ofready-to-use conjugate into the wells. Leave one well empty for the substrateblank. 6) Coverplate with the enclosed foil and incubate at room temperature for 30 minutes. 7) Emptythe wells of the plate (dump or aspirate) and add 300 uL ofdiluted washing solution. This procedure is repeated totally three times.Rests of the washing buffer are afterwards removed by gentle tapping of themicrotiter plate on a tissue cloth. 8) Pipet100 uL each ofthe ready-to-use substrate into the wells. This time also the substrate blankis pipetted. 9) Coverplate with the enclosed foil and incubate at room temperature for 20 minutesin the dark(e.g. drawer). 10) Toterminate the substrate reaction, pipet 100 uL each ofthe ready-to-use stop solution into the wells. Pipet also the substrateblank. 11) Afterthorough mixing and wiping the bottom of the plate, perform the reading ofthe absorption at 450 nm (optionally reference wavelength of 620 nm). Thecolor is stable for at least 60 minutes.