Description

·         Used antigen: LKM-1, LC-1, SLA/LP, Sp100, gp210, ASGPR, PML, Nup62, M2, 3E(BPO), OGDC-E2, PDC-E2 and Ro52.

·         Antigens are applied in the form of microdots (spots) in triplets to a nitrocellulose membrane in a microtiter well format.

·         If specific antibodies are present in the sample, they will bind to the respective separate antigens.

·         The complex is labelled with Conjugate and detected through a colour reaction with Substrate (BCIP/NBT).

·         The kit enables 48 tests.

·         Short incubation periods, total assay time: approximately 1.5 h.

·         Each individual test includes control spots, which serves to check functionality of the kits, and also the spots for checking the presence of the conjugate.

·         Included positive controls enable validation of the tests.

·         For quantitative evaluation the wells contain calibration spots.

·         All reagents are supplied ready to use.

·         The kit may be used sequentially for smaller batches of samples (reagents are provided in sufficient quantities).

·         Evaluation using Microblot-Array reader and the appropriate software.

Applications

·         Confirmation for ELIA or other screening tests.

·         Identification of individual antibody types, differentiation of individual autoimmune liver diseases.

Brief assay procedure

1.       Dilute samples of serum/plasma (1:51)

2.       Pipette diluted samples

3.       Incubate at room temperature for 30 min.

4.       Wash 3 times for 5 min.

5.       Add Conjugate

6.       Incubate at room temperature for 30 min.

7.       Wash 3 times for 5 min.

8.       Add Substrate (BCIP/NPT)

9.       Incubate at room temperature for 15 min.

10.   Wash 2 times for 5 min. in distilled water

11.   Dry and evaluate test