FLOT1/2 IIFT in MS patients kit (EUROIMMUN) samples 101-123

OBJECTIVES

To determine the existence of anti-FLOT1/2 antibodies in a subset of MS patients.

SAMPLES

1. 101
2. 102
3. 103
4. 104
5. 105
6. 106
7. 107
8. 108
9. 109
10. 110
11. 111
12. 112
13. 113
14. 114
15. 115
16. 116
17. 117
18. 118
19. 119
20. 120
21. 121
22. 122
23. 123
24. Negative control (provided in the kit)
25. Positive control (provided in the kit)

MATERIALS

• Materials from the EUROIMMUN kit FA 1124-####-50 Anti-Flotilin (FLOT1/2) IIFT, including:
  • 5 BIOCHIPs containing FLOT1/2-transfected HEKs
  • PBS-Tween (Tween at 2ml/l)
  • Positive and negative controls.
  • Fluorescein-labelled anti-human globulin antibodies.
  • Glycerol mounting medium.
  • Cover glasses for the BIOCHIPs.
• Reagent Tray

PROCEDURE

• Serum samples were diluted 1:10 in PBS-Tween.
• 30 microL of the serum dilution were added to each reaction field in the reagent tray, and the BIOCHIP slides were placed on them.
• The reaction was incubated for 30 minutes at RT.
• The BIOCHIPS were rinsed with a flush of PBS-Tween and then immersed in a cuvette containing PBS-Tween for 5 minutes while placed in a rotary shaker.
• While the slides were being cleaned, the reaction tray was also cleaned with distilled water and prepared for the next reaction by placing 25 microL drops of the fluorescein-labelled antibody in each reaction field. The BIOCHIPs were then dried on the back and sides and immediately placed on the reaction tray.
• The reaction was incubated or 30 minutes at RT.
• The BIOCHIPS were rinsed with a flush of PBS-Tween and then immersed in a cuvette containing PBS-Tween (changed since the last wash) for 5 minutes while placed in a rotary shaker.
• The mounting medium was placed on the cover glasses while lined up on the base of the reaction tray. When the wash was finished, the BIOCHIP slides were cleaned on the back and sides and immediately placed on the cover glasses.
• The BIOCHIPS were then read in the fluorescence microscope.

RESULTS

1. Negative.
2. Negative.
3. Not visible.
4. Not visible.
5. Not visible.
6. Negative.
7. Negative.
8. Negative.
9. Negative.
10. Negative.
11. Negative.
12. Negative.
13. Negative.
14. Negative.
15. Negative.
16. Negative.
17. Negative.
18. Negative.
    
19. Negative.
    
20. Negative.
    
21. Negative.
    
22. Negative.
23. Negative.
    
24. Negative.
25. Positive.

HEKs from the first BIOCHIP (samples 1-5) were smeared. 1 and 2 were partially visible but 3 to 5 were too smeared to maked a determination and will be repeated in the next batch.