What is Sandwich Chemiluminescence immunoassay

1. Add 100 ul of 1ug/ml capture antibody or cell lysate or serum in the microwell strips. Seal with tape and press firmly onto top of microwells. Incubate the plate for 2 hr at room temperature. Alternatively, the plate can be incubated overnight at 4°C.

2. Gently remove the tape and wash wells:

(1) Discard plate contents into a receptacle.

(2) Wash 4 times with 1X Wash Buffer, 150 µl each time per well.

(3) For each wash, strike plates on fresh paper towels hard enough to remove the residual solution in each well, but do not allow wells to dry completely at any time

(4) Clean the underside of all wells with a lint-free tissue.

3. Add 50 µl of antigen or analyte need to detect to each well. Seal with tape and incubate the plate at room temperature for 1 hr.

4. Repeat wash procedure (Step 2).

5. Add 50 µl of HRP-linked secondary antibody to each well. Seal with tape and incubate the plate at room temperature for 30 min.

6. Repeat wash procedure (Step 2).

7. Prepare detection reagent working solution by mixing equal parts 2X luminol Reagent and 2X Peroxide.

8. Add 50 µl of the detection reagent working solution to each well.

Use a plate-based luminometer set at 425 nm to measure Relative Light Units (RLU) within 1–10 min following addition of the substrate. (Optimal signal intensity is achieved when read within 10 min.)

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Picture loading failed. Guidence of GeneMedi's protocol / procedure for the diagnostics application:
         1. CLIA
         2. ELISA: (1) Direct ELISA (2) Indirect ELISA (3) Competitive ELISA (4) Sandwich ELISA
         3. LFIA: (1) Sandwich format (2) Competitive format (3) Multiplexed lateral flow assays
         4. PETIA
         5. Immunonephelometry
         6. IHC
         7. FACS: (1) Antibody titration protocol using 96 well plates (2) Protocol for cell sorting
         8. Octet system
   Summary of the advantages and disadvantages of the different immunoassay

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