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P4653

Sigma-Aldrich

Phosphatase, Alkaline from bovine kidney

lyophilized powder, ≥5 units/mg protein (Activity determined in diethanolamine buffer, pH 9.8)

Synonym(s):
Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
eCl@ss:
42010105

form

lyophilized powder

specific activity

≥5 units/mg protein (Activity determined in diethanolamine buffer, pH 9.8)

composition

Protein, ~35% biuret

storage temp.

−20°C

Application

Alkaline phosphatase is used for conjugation to antibodies and other proteins for ELISA, Western blotting, and histochemical detection. It is routinely used to dephosphorylate proteins, such as casein, and nucleic acids[1]. It may be used for protein labeling when high sensitivity is required. Alkaline phosphatase may be also be used to dephosphorylate the 5′-termini of DNA or RNA to prevent self-ligation. DNA or RNA can also be tagged with radiolabeled phosphate (via T4 polynucleotide kinase) after dephosphorylation with alkaline phosphatase.. Product P4653 is from bovine kidney and is provided as a lyophilized powder containing Tris-citrate buffer salts.

Biochem/physiol Actions

Alkaline phosphatase dephosphorylates proteins and nucleic acids.

Unit Definition

One unit will hydrolyze 1 μmole of 4-nitrophenyl phosphate per minute at pH 9.8 at 37 °C. Previously sold in "glycine" units.

Physical form

Lyophilized powder containing Tris-citrate buffer salts

Analysis Note

Assays in Tris buffer, pH 10.2 give activity approx. 2X that in the glycine buffer system. Assays in diethanolamine buffer, pH 9.8 give activity approx. 5X that of the glycine buffer system.
Protein determined by biuret

Certificate of Analysis

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Certificate of Origin

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Technical Information

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