DiaPharma Group, Inc.

DISCONTINUED

For in vitro diagnostic use

Direct EIA (enzyme immunoassay) test kit for the detection and measurement of activated factor XII.

Kit Components

1. Anti-FXIIa conjugate: 1x11 ml vial containing alkaline phosphatase-labeled sheep anti-human activated FXII antibody.
2. Substrate: 1x15 ml vial containing Mg2+ as enzyme cofactor and phenolphthalein monophosphate (PMP) in buffer solution.
3. Stop Solution: 1x15 ml vial containing sodium hydroxide and EDTA as a chelating agent in carbonate buffer pH>10.
4. FXIIa wash buffer concentrate: 2x20 ml vials containing borate buffer. Supplied x20 Conc.
5. FXIIa coated wells: 6x16 well microtitre strips coated with anti-FXIIa monoclonal antibody.
6. FXIIa standards: 1.0 ml each of 5 standards containing purified FXIIa (b-XIIa) in buffer.
7. FXIIa controls 1 and 2: 2x0.5 ml each containing freeze-dried human plasma with FXIIa.
8. Pack leaflet.
9. Microstrip holder.

Specimen Collection and Storage

Venipuncture should be clean and deliberate to avoid activation of coagulation.

Collect blood in 0.109M trisodium citrate anticoagulant, using the ratio 1 volume of anticoagulant to 9 volumes of blood. The use of other anticoagulants has not been validated.

Whole blood should be centrifuged at 3000 x g for 10 minutes. Plasma must be separated within 1 hour of sample collection.

Do not store samples at 2-8°C as this may initiate cold activation in some samples. Plasma samples can be stored for up to three hours at room temperature prior to assay.

Expected Values

(references available on request)

Imprecision

Summary of the Protocol

1. Reconstitute the controls.
2. Pipette 100 ml of standards, controls and patient samples in appropriate wells.
3. Incubate at 18-25°C for 60±10 minutes.
4. Wash strips x5.
5. Add 100 ml of AFT conjugate to each well.
6. Incubate at 18-25°C for 60±+5 minutes.
7. Wash strips x5.
8. Add 100 ml of substrate to each well.
9. Incubate for at 18-25°C for 15±2 minutes.
10. Add 100 ml of stop solution to each well.
11. Read absorbance at 550 nm.