2013 Promega catalogue

Cell Signaling
Alkaline Phosphatases
Alkaline Phosphatase, Calf Intestinal (CIAP)
Product Size Conc. Cat.# Price ($)
Alkaline Phosphatase, Calf Intestinal 1,000 u 1 u/µl M1821 107
Alkaline Phosphatase, Calf Intestinal (HC) 1,000 u 20 u/µl M2825 107
Available Separately Size Cat.# Price ($)
CIAP Buffer Pack 1.5 ml M1833 32
For Laboratory Use.
Description: Alkaline Phosphatase, Calf Intestinal (CIAP), catalyzes the
hydrolysis of 5´-phosphate groups from DNA, RNA, and ribo- and deoxyribonucleoside
triphosphates. This enzyme is used to prevent recircularization
and religation of linearized cloning vector DNA by removing phosphate groups
from both 5´-termini and may also be used for the dephosphorylation of 5´
phosphorylated ends of DNA or RNA for subsequent labeling with [ 32 P]ATP and
T4 Polynucleotide Kinase. CIAP is active on 5´ overhangs, 5´ recessed and blunt
ends.
Features:
• Available at High Concentration: Cat.# M2825 contains 1,000 units of
CIAP at 20u/μl.
• Blue/White Cloning Qualified: Promega’s blue/white cloning assay
provides a higher level of quality control for enzymes used in cloning applications.
• Provided with 10X Reaction Buffer: 0.5M Tris-HCl (pH 9.3 at 25°C),
10mM MgCl 2 , 1mM ZnCl 2 , 10mM spermidine.
Storage Conditions: Store at –20°C.
Protocol
Alkaline Phosphatase, Calf Intestinal (CIAP) Protocol
Part#
9PIM182
TSAP Thermosensitive Alkaline Phosphatase
Product Size Cat.# Price ($)
TSAP Thermosensitive Alkaline Phosphatase 100 units M9910 113
For Laboratory Use.
Description:
TSAP Thermosensitive Alkaline Phosphatase catalyzes the removal of 5´ phosphate
groups from DNA, thus preventing the recircularization and religation of
linearized cloning vector DNA during ligation. It is effective on 3´ overhangs, 5´
overhangs and blunt ends. It is also useful for preparing DNA for 5´ end-labeling
by removing existing phosphate groups from the 5´ end.
TSAP is irreversibly inactivated by heating at 74°C for 15 minutes.
Therefore, a DNA cleanup step is not required before proceeding to a ligation
reaction. TSAP is fully active in all restriction enzyme reaction buffers tested
under the conditions listed below, facilitating a streamlined restriction digestion,
dephosphorylation and ligation reaction.
Features:
• Easy To Use: TSAP is active in all Promega restriction enzyme buffers,
eliminating any cleanup steps or buffer swaps.
• Convenient: TSAP is irreversibly inactivated by heating at 74°C for 15
minutes. This allows streamlining of the restriction enzyme digestion, dephosphorylation
and ligation procedure by eliminating the need for cleanup
after alkaline phosphatase treatment.
• Blue/White Cloning-Qualified: Promega’s blue/white cloning assay
provides a higher level of quality control for enzymes used in cloning applications.
• Provided with Promega MULTI-CORE Buffer.
Storage Conditions: Store at –20°C. See the expiration date on the label.
Protocol
TSAP Thermosensitive Alkaline Phosphatase Product Information
Part#
9PIM991
5
Cloning and DNA Markers
Comparison of Alkaline Phosphatases.
TSAP CIAP*
Heat Inactivated Yes No
Inactivation Temperature 74 N/A
Incubation Time 15 min 2 × 30 min
Special Buffer Required/Recommended No Yes
Active in all Promega
Restriction Enzyme Buffers Yes No
Units required in different RE Buffers 1–2 N/A
Blue/White Cloning-Qualified Yes Yes
Only TSAP does not require a special buffer and is active in all Promega restriction
enzyme buffers, making it the most convenient and cost-effective choice.
CIAP = Calf Intestinal Alkaline Phosphatase
9493LA
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