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DÍEZ MONTORO, R. & COL. AN. R. ACAD. NAC. FARM.
3.2. Reagents
DM = Solution of 125I-labelled Insulin in a protein-based buffer.
Estimated concentration ˜ 50 pmol/L.
PT = Plastic tubes with rabbit anti-Insulin immunoglobulin immo-
bilized to the inside wall.
DQ = Insulin standard solutions.
These reagents were included in the Insulin RIA DPC kit.
GL = Glycerol Merck pro analysis.
DS = Solution of NaCl 2.05 M.
Several tube series were prepared as per the Table I.
Table I. Preparation of Coated Tubes Containing 125I-Insulin
PT 1-6 7-12 13-18 19-100
DM (mL) 0.25 0.50 0.75 1
H20 (mL) 0.75 0.50 0.25 0
They were left to react overnight. The next day, they were decant-
ed and washed with 2 mL distilled water.
Solutions were prepared as per Table II.
Table II. Preparation of Solutions Containing Insulin, Glycerol and NaCl
Solution 1 2 3 4 5 6 7 8 9 10
DQ (µL) 25 50 75 100 100 100 100 100 100 800
GL (mL) 0 0 0 1 2 3 0 0 0 0
DS (µL) 100 100 100 100 100 100 200 300 400 800
H2O (mL) 7.875 7.850 7.825 6.8 5.8 4.8 7.7 7.6 7.5 62.4
q (pmol/L) 7.84 15.7 23.5 31.4 31.4 31.4 31.4 31.4 31.4 31.4
q = Concentration of Insulin
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