Elisa

1. 10X Adsorption buffer: 1 M sodium bicarbonate, pH 9.6 (adjust with NaOH).

2. Peptide solution/suspension at 1 mg/mL in phosphate-buffered saline (PBS) stored at -20°C (see Note 4).

3. Antipeptide antibody.

4. "Immulon 2" high binding capacity 96-well microtiter plates (Dynatech).

5. Donkey anti-rabbit Ig-horseradish peroxidase conjugate (DaRIg-HRP).

6. Swine anti-rabbit Ig-horseradish peroxidase conjugate (SwaRIg-HRP) (APBiotech or DAKO).

7. Tris-buffered saline (TBS): 25 mM Tris-HCl/144 mM NaCl, pH 8.1.

8. Blocking buffer (TM): TBS containing 2% dried milk powder (Marvel).

9. Assay buffer (TMT): TBS containing 2% dried milk powder and 0.2% Tween-20.

10. Substrate solution for HRP: 1-mM ABTS (2,2' azinobis [3-ethylbenzthiazoline-6 sulphonic acid]) (Sigma) in 0.1 M sodium acetate pH 5.0. Add 1 ^L of 30% hydrogen peroxide per 1 mL of ABTS solution just before use. Discard any old ABTS stocks that have a noticeable green color when dissolved in the absence of hydrogen peroxide.

11. Stop solution for peroxidase assays: 5% sodium dodecyl sulfate (SDS).

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