· The antigens of Toxoplasma gondii are separated by gel electrophoresis, and then transferred to the nitrocellulose membrane (i.e. to the respective WB strips).
· If specific antibodies are present in the sample, they will bind to the respective antigens Mr = 30 kDa.
· The complex is labelled with Conjugate and detected through a colour reaction with substrate (BCIP/NBT).
· The kit enables 16 tests.
· High sensitivity and specificity.
· Total assay time: approximately 3 h.
· For test validity verification, the strips are equipped with a control band (which indicates the kit functionality and sensitivity).
· Positive and Negative controls included enabling validation of the tests.
· Identical (i.e. interchangeable) reagents (except for Conjugate) in all BLOT kits.
· All reagents supplied are ready to use.
· The kit may be used sequentially for smaller batches of samples, reagents are provided in sufficient quantities.
· Evaluation of results – the intensity of the bands can be evaluated visually or by means of software (Immunoblot Software).
· Laboratory test for the detection of infection with Toxoplasma gondii in humans, confirmatory test to ELISA.
Brief assay procedure:
1. Dilute controls and tested samples (1:51).
2. Pipette diluted controls and samples into the channels of the tray and insert the WB strips.
3. Incubate for 60 minutes at room temperature using a shaking apparatus.
4. Wash 3 times for 5 minutes.
5. Add Conjugate.
6. Incubate for 60 minutes at room temperature using a shaking apparatus.
7. Wash 3 times for 5 minutes.
8. Add substrate (BCIP/NBT).
9. Incubate for 10 minutes at room temperature using a shaking apparatus.
10. Wash 2 times for 5 minutes in distilled water.
11. Dry the strips and evaluate results using the validation strip enclosed.