Alkaline Phosphatase, Calf Intestinal (CIP) Catalog # Size Concentration M0290S   STOCK! 1,000 units 10,000 units/ml M0290L 5,000 units 10,000 units/ml Download: Technical Bulletin| Removes 5´ phosphates from DNA, RNA, rNTPs and dNTPs Prevents recircularization of cloning vectors Dephosphorylates serine, threonine and tyrosine residues in proteins Supplied with 10X Reaction Buffer Description: Alkaline Phosphatase catalyzes the removal of 5´ phosphate groups from DNA, RNA, ribo- and deoxyribonucleoside triphosphates. Since CIP-treated fragments lack the 5´ phosphoryl termini required by ligases, they cannot self-ligate (1). This property can be used to decrease the vector background in cloning strategies. Source: Calf intestinal mucosa Applications: Removing 5´ and 3´ phosphoryl groups from nucleic acids Preparing templates for 5´end labeling Preventing fragments from self ligating Dephosphorylation of proteins Reagents Supplied: NEBuffer 3 (10X) Enzyme Properties Heat Inactivation: No Molecular Weight: Apparent: 138 kDa Specific Activity: 3,500 units/mg Reaction & Storage Conditions Reaction Conditions: 1X NEBuffer 3 Incubate at 37°C. 1X NEBuffer 3: 50 mM Tris-HCl 100 mM NaCl 10 mM MgCl2 1 mM Dithiothreitol pH 7.9 @ 25°C Unit Definition: One unit is defined as the amount of enzyme that hydrolyzes 1 µmol of p-nitrophenylphosphate to p-nitrophenol in a total reaction volume of 1 ml in 1 minute at 37°C (2) in 1 M diethanolamine-HCl (pH 9.8) with 0.5 mM MgCl2 and 10 mM p-nitrophenylphosphate. These conditions are only used for quantitating enzyme activity. Concentration: 10,000 units/ml Storage Conditions: 10 mM Tris-HCl 50 mM KCl 1 mM MgCl2 0.1 mM ZnCl2 50% Glycerol pH 8.2 @ 25°C Storage Temperature: -20°C Notes General notes: CIP can be used in NEBuffers 2, 3 or 4 as well as the Unique NEBuffers for EcoR I and BamH I. NEBuffer 3 gives optimum activity.