AP Magenta 3 Component Membrane Substrate

Store at temperatures between 2-8ºC.

Stable for a minimum of 18 months from the manufactured date when properly stored between 2-8ºC.

Leinco’s AP Magenta 3 Component Membrane Substrate is a precipitating substrate used with the enzyme alkaline phosphatase designed for membrane blotting procedures. The TAC Buffer coupled with a substitute, naphthol, is enzymatically hydrolyzed. In the presence of hexazotized substrate, a fine magenta precipitate is produced at the sites of alkaline phosphatase activity.
Components:
1. TAC Buffer (Modified Gormori’s Tris Azo-Coupling Buffer) - 4 x 100 ml or 1 x 1 L
2. Initiator (Hexa-azonium salt initiator) - 1 x 8 ml or 1 x 20 ml
3. Magenta Solution - 1 x 8 ml or 1 x 20 ml

1. Add 2 drops, or 100 µl, of Magenta Solution into an appropriate sized test tube or collection vessel.
2. Add 2 drops, or 100 µl, of Initiator Solution to the 100 µl of Magenta Solution in step one. Gently mix and allow solution to sit for 3 to 5 minutes.
3. After the 3 to 5 minute incubation, add 5 ml of TAC Buffer and mix. USE IMMEDIATELY! (NOTE: Reagent is stable for up to one hour. Some color and turbidity may develop over this time frame. However, this does not affect product performance.)
4. Apply sufficient substrate solution to cover the membrane completely.
5. Incubate samples 10 minutes to 60 minutes, at room temperature, depending on individual preferences of staining intensity.

NOTE: Sites of enzymatic activity vary from pink to magenta depending in antigenicity and length of staining. Dilution of substrate is not recommended. To reduce the intensity of the reaction, it is recommended that antibodies or conjugates be diluted.