Alkaline Phosphatase 2-Component System - Code E552

Directions for Use

This system contains Alkaline Phosphatase at a 10 mg/ml (30,000 units/ml) concentration along with a second tube, containing a dilution/storage buffer.  This concentration is suitable for conjugation to secondary antibodies or cellular components such as biotin or avidin.  For DNA dephosphorylation, it is recommended that the enzyme be diluted as follows:

Add the entire contents of the tube of enzyme to the included dilution buffer; pipette briefly to mix.  The dilution buffer may be used to rinse the tube of enzyme to remove any remaining enzyme residue.

Concentration of the diluted enzyme is 3 units/l.  For most applications, 0.5 l of diluted enzyme will dephosphorylate up to 20 pmol of 5' ends in a 50 l reaction volume @ 37C.  The enzyme at this concentration is stable for up to one year when stored at 4C.

Enzyme storage/dilution buffer:

50% Glycerol

1mM MgCl2

0.1mM ZnCl2

pH 7.5

Reaction conditions will vary, according to specific application.  Normal buffer conditions for DNA dephosphorylation are:

20mM TrisCl, pH 8.0

1mM MgCl2

1mM ZnCl2

1 to 20 pmol 5' ends

0.5 l diluted Alkaline Phosphatase

tel (33) (0) 4 70 03 88 55 

fax(33)(0) 4 70 03 82 60

 http://www.interchim.com

Vos contacts:

Etienne Boireau - Olivier Cadas - Olivier Alméras

hot line  33 (0) 4 70 03 73 06