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P4978

Sigma-Aldrich

Phosphatase, Alkaline from calf intestine

buffered aqueous glycerol solution

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Synonym(s):
CIAP, CIP, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
CAS Number:
Enzyme Commission number:
MDL number:
eCl@ss:
42010105
NACRES:
NA.53

grade

for 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

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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.

Components

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.

Unit Definition

One unit will hydrolyze 1 μmole of p-nitrophenyl phosphate per min at 37 °C.

Other Notes

Alkaline phosphatase in 5 mM EDTA (pH 8) will be irreversibly heat inactivated at 75 °C for 10 minutes.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Christopher J Moore et al.
Genes & development, 35(3-4), 286-299 (2021-01-16)
RNase E is an essential, multifunctional ribonuclease encoded in E. coli by the rne gene. Structural analysis indicates that the ribonucleolytic activity of this enzyme is conferred by rne-encoded polypeptide chains that (1) dimerize to form a catalytic site at
Yusheng Zhao et al.
Genes & development, 35(11-12), 888-898 (2021-05-15)
Plants monitor many aspects of their fluctuating environments to help align their development with seasons. Molecular understanding of how noisy temperature cues are registered has emerged from dissection of vernalization in Arabidopsis, which involves a multiphase cold-dependent silencing of the
J Juhásová et al.
Physiological research, 60(3), 559-571 (2011-03-16)
Mesenchymal stem cells (MSCs) have been repeatedly shown to be able to repair bone defects. The aim of this study was to characterize the osteogenic differentiation of miniature pig MSCs and markers of this differentiation in vitro. Flow-cytometrically characterized MSCs
A D Bakker et al.
Journal of dental research, 93(4), 394-399 (2014-02-05)
Mechanosensitive osteocytes regulate bone mass in adults. Interleukin 6 (IL-6), such as present during orthodontic tooth movement, also strongly affects bone mass, but little is known about the effect of IL-6 on osteocyte function. Therefore we aimed to determine in
M D McKee et al.
Journal of dental research, 92(8), 721-727 (2013-05-23)
Phosphatases are involved in bone and tooth mineralization, but their mechanisms of action are not completely understood. Tissue-nonspecific alkaline phosphatase (TNAP, ALPL) regulates inhibitory extracellular pyrophosphate through its pyrophosphatase activity to control mineral propagation in the matrix; mice without TNAP

Protocols

CIP is used to remove 5’-phosphate groups from DNA, RNA and both ribo and deoxy-ribonucleoside triphosphates. Detailed protocol on how to dephosphorylate DNA.

Enzymatic Assay of Alkaline Phosphatase, Diethanolamine Assay (EC 3. 1. 3. 1)

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