Alkaline Phosphatase-conjugate

Western Blot Analysis of Epitoped-tagged Proteins Using The Chemifluorescent Detection Method - for alkaline phosphatase conjugated antibodies

Secondary antibody: Goat anti-mouse alkaline phosphatase conjugated - Biorad #170-6520. Substrate: ECF chemifluorescent substrate - Amersham #RPN5785. Mix substrate with accompanying buffer as per manufacturer’s

（共享）ELISA原理与实验方法

, incubate for 30 min at 37°C. ( 5g powdered milk in 95 ml PBS)5 Rinse with PBS 3x.6 Add 50 µl of antibody, incubate 30 min at 37°C.7 Rinse with PBS 3x.8 Add 50 µl of goat anti-mouse IgG conjugate with alkaline phosphatase.(1 µl of conjugate in 4 ml of PBS

Purification of mAb (IgG)

buffer (pH9.0) ~ 200 ml. Filter protein mixture Add protein mixture or culture supernatant containing Ab (adjust pH to 7.8 with Binding buffer ; red color) to the Protein A column. Mouse antibodies of the IgG1 subclass