P4978
Phosphatase, Alkaline from calf intestine
buffered aqueous glycerol solution
Synonym(s):
CIAP, CIP, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
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About This Item
CAS Number:
UNSPSC Code:
12352204
eCl@ss:
42010105
NACRES:
NA.53
MDL number:
Concentration:
≥10,000 units/mL
grade
Molecular Biology
form
buffered aqueous glycerol solution
mol wt
~80 kDa
concentration
≥10,000 units/mL
UniProt accession no.
foreign activity
DNase, RNase, none detected
shipped in
wet ice
storage temp.
−20°C
Gene Information
cow ... ALPI(280993)
General description
Alkaline phosphatase is a phosphomonoesterase that catalyzes the cleavage of terminal phosphates from nucleic acids. The free 5′-OH can be phosphorylated with polynucleotide kinase and γ32P-ATP to produce γ32P-end-labeled nucleic acids. Linearized cloning vectors can be prevented from recircularizing by dephosphorylation with alkaline phosphatase.
Application
Commonly used to remove the 5′-terminal phosphate from nucleic acids during molecular cloning reactions.
Other Notes
Alkaline phosphatase in 5 mM EDTA (pH 8) will be irreversibly heat inactivated at 75 °C for 10 minutes.
Alkaline phosphatase is provided in a solution of 10 mM Tris-HCl (pH 8.2), 50 mM KCl, 1 mM MgCl2, and 0.1 mM ZnCl2, in 50% (w/v) glycerol. 10X CIP Buffer, Product Number C3225, is included.
One unit will hydrolyze 1 μmole of p-nitrophenyl phosphate per min at 37 °C.
Storage Class
10 - Combustible liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
Regulatory Information
低风险生物材料
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Protocols
碱性磷酸酶的酶法测定，二乙醇胺法（EC 3.1.3.1)
CIP可用于脱去DNA、RNA及核糖和脱氧核糖核苷三磷酸上的5’-磷酸基团。如何进行DNA去磷酸化的详细实验方案。
Enzymatic Assay of Alkaline Phosphatase, Diethanolamine Assay (EC 3. 1. 3. 1)