Analysis Note

The activity at pH 8.0 at 25 °C is approximately the same as at pH 10.4 at 37 °C.

Application

Alkaline phosphatase is used for conjugation to antibodies and other proteins for ELISA, Western blotting, and histochemical detection. It may be used for protein labeling when high sensitivity is required. Product P4252 is an ammonium sulfate suspension in glycine buffer.

Alkaline phosphatase is used for conjugation to antibodies and other proteins for ELISA, Western blotting, and histochemical detection. It may be used for protein labeling when high sensitivity is required. Product P4252 has been used for bulk digestion of tRNA . Product P4252 is provided in the ratio of 30-90 units/mg of protein.

Biochem/physiol Actions

Alkaline phosphatase, from Escherichia coli, is a dimeric, non-glycosylated protein which mainly reside in the periplasmic space. Three known isoforms exist. The enzyme requires zinc, and is activated by magnesium. E. coli akaline phosphatase has a broad specificity for phosphate esters.

Alkaline phosphatase, from Escherichia coli, is a dimeric, non-glycosylated protein which mainly reside in the periplasmic space. Three known isoforms exist. The enzyme requires zinc, and is activated by magnesium. E. coli akaline phosphatase has a broad specificity for phosphate esters.

Caution

Presence of 0.1% SDS in the alkaline phosphatase assays will inhibit any nuclease contamination in the sample.

Packaging

100, 200, 1000 units in glass bottle

Physical form

Suspension in 2.5 M (NH₄)₂SO₄

Unit Definition

One unit will hydrolyze 1.0 µmole of p-nitrophenyl phosphate per min at pH 10.4 at 37 °C.