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P4978

Sigma-Aldrich

Phosphatase, Alkaline from calf intestine

buffered aqueous glycerol solution

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

Quality Level

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

Certificate of Analysis

Certificate of Origin

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...
Timothy M Chlon et al.
Journal of virology, 88(19), 11315-11326 (2014-07-18)
DNA repair plays a crucial role in embryonic and somatic stem cell biology and cell reprogramming. The Fanconi anemia (FA) pathway, which promotes error-free repair of DNA double-strand breaks, is required for somatic cell reprogramming to induced pluripotent stem cells...
Jong Kil Lee et al.
The Journal of experimental medicine, 211(8), 1551-1570 (2014-07-23)
In Alzheimer's disease (AD), abnormal sphingolipid metabolism has been reported, although the pathogenic consequences of these changes have not been fully characterized. We show that acid sphingomyelinase (ASM) is increased in fibroblasts, brain, and/or plasma from patients with AD and...
Peter M Krawitz et al.
American journal of human genetics, 92(4), 584-589 (2013-04-09)
Recently, mutations in genes involved in the biosynthesis of the glycosylphosphatidylinositol (GPI) anchor have been identified in a new subclass of congenital disorders of glycosylation (CDGs) with a distinct spectrum of clinical features. To date, mutations have been identified in...

Protocols

Calf Intestinal (CIP) Alkaline Phosphatase for Nucleic Acid Dephosphorylation

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.

Diethanolamine Assay for the Enzymatic Assay of Alkaline Phosphatase (EC 3.1.3.1)

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

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