DiaPharma Group, Inc.

For research use only. Not for use in diagnostic procedures.

Intended Use

Biopool TintElize® D-dimer is intended for quantitative determination of human D-dimer in plasma by enzyme immunoassay.

Principle

The Biopool TintElize® D-dimer utilises the double antibody principle. Plasma sample or standard containing D-dimer is added to a microtest well, which is coated with a monoclonal antibody, MA-8D3, against D-dimer. After an incubation sufficient to allow >85 % of the D-dimer to bind to the coat antibodies, HRP labelled Fab fragments of anti D-dimer IgG are added. These are allowed to react with the adsorbed Ddimer. The wells are emptied and washed to remove unbound conjugate after which peroxidase substrate (OPD/H2O2) is added. The quantity of yellow colour developed is directly proportional to the amount of D-dimer present in the sample.

Reagents -- Composition, Packaging, Storage and Stability

The unopened reagents should be stored at 2-8°C and be used prior to the expiration date. Reconstituted reagents are stable for two weeks if stored as recommended except the standards which are stable for two days if stored as recommended.

1. Microtest strips,12 pcs: Framed 8-well strips, pre-coated, with monoclonal antibody MA-8D3, and prefilled with non-immune mouse IgG and indicator dye.
2. Assay Buffer 15X, 2 vials: Potassium phosphate, Borate, EDTA, KCl, and Tween 20 buffer solution, pH 7.5 sufficient to make 2 x 0.5 L solution.
3. Standard 0 ng/ml, 3 x 0.5 ml: Lyophilised, citrated human plasma depleted of D-dimer.
4. Standard 1000 ng/ml, 3 x 0.5 ml: Lyophilised, citrated human plasma enriched with D-dimer.
5. Conjugate, 6 ml: HRP labelled Fab fragments of anti D-dimer antibody.
6. Substrate, 2 ml: Lyophilised ortho-phenylenediamine (OPD) with buffer salts.
7. Hydrogen Peroxide, 2 ml: 0.15 % H2O2 in water.
8. Reagent Reservoirs, 6 Disposable cardboard trays

Material required but not provided

• Pipette 8-channel or repeating for 25-100 microliters
• Pipettes 1-channel covering 25-1000 microliters
• Squeeze bottle
• Paper towels or thin sponge
• Small plastic tubes (2-5 ml)
• Purified water (distilled or deionized and sterile filtered)
• Sulphuric acid H2SO4 1.6 mol/L
• Microtest plate spectrophotometer operable at 492 nm
• Microtest plate shaker with an orbital of 3 mm.