Bio-Rad Mouse Typer Isotyping Kit - Manual (Page 5)

7. 96-well polystyrene microplates (Bio-Rad catalog number
224-0096).
8. Pipet tips (Bio-Rad catalog number 223-9302, nonsterile).
9. Octapette ® pipet, 100 ml (Bio-Rad catalog number 224-4800).
10. Reagent reservoirs (Bio-Rad catalog number 224-4872).
11. Automatic ELISA plate reader (Bio-Rad Model 3550 Micro-
plate Reader, catalog number 170-6601 or Model 3550-UV
Microplate Reader, catalog number 170-6638, or Model 550
Microplate Reader, catalog number 170-6750).
Section 2
Sub-Isotyping Assay
2.1 Solutions
The solution volumes that follow are recommended for an
ELISA assay typing of 10 mouse monoclonal antibodies in one
microtitration plate.
Phosphate buffered (0.01 M phosphate buffer, pH 7.2)
saline, PBS, 300 ml Add 0.105 g sodium monobasic phosphate,
0.600 g sodium dibasic phosphate, and
2.550 g sodium chloride to 250 ml distilled,
deionized water. Adjust to pH 7.2 with HCl
and bring to 300 ml with water.
Phosphate buffered Divide the above solution into 100 and
saline wash solution, 200 ml fractions. Add 0.10 ml Tween 20
PBS-Tween, 200 ml
to the 200 ml fraction.
Antigen solution, Dissolve or dilute antigen preparation in
10 ml 10 ml PBS to a final concentration of 1-
10 µg/ml.
3
LIT78D 8/7/98 04:08 PM Page 3