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Coated Plate Immunoassay on the MicroBeta - eg. TSH - IRMA

0.9% NaCl add water 90 ml

2. Add anti-TSH into coating buffer (antibody concentration 1.0 μg/200μL/well)

3. Dispense 200 μL/well

4. Incubate overnight in a moist chamber

5. Wash 3 x with the wash solution

6. Saturate 1 hour at room temperature with 200 μL of 50 mM TSA-buffer containing 0.5% BSA
Saturation Buffer formulation:
50 mM Tris = 0.606 g Tris (Sigma)
0.9% NaCl = 0.9 g
conc HCl 0.30 mL
7.5% BSA 6.66 mL
add 100 mL, pH 7.77

7. Aspirate to dryness

8. Cover with re-sealable seal and store at 4C

**ASSAY PROCEDURE OF TSH

Previously coated plates. Farmos 125I approx. 450 kBq/11 ml anti-TSH.

ASSAY PROCEDURE

1. Add 100 μL standard and samples in duplicates to the plates

2. Add 100 μL tracer

3. Incubate at room temperature for 2 hours (or overnight)

4. Wash 5 x with wash solution with a suitable plate washer

Wash solution formulation:
Tris-HCl 0.005 M (5 mM), pH 7.5
0.9% NaCl
0.005% Tween-20

5. Decant excess wash solution (& dry if using Meltilex)

6. Add scintillant to each well, shake for 5 minutes on a plate shaker

7. Count in MicroBeta


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