Alkaline Phosphatase, Calf Intestine (Endonuclease, Exonuclease and RNase Free)

Molecular Biology / MB-Enzymes, Phosphatase

-20°C

Supplied as a lyophilized powder. No preservative added. Reconstitute with sterile ddH2O.

Alkaline phosphatases (APs) are highly ubiquitous enzymes, present in all species from bacteria to man. In humans, APs are encoded by a multi-gene family composed of four loci; i.e., tissue- nonspecific AP, also called bone/liver/kidney AP, (Weiss et al, J. Biol. Chem., 264, 12002-12010, 1988), intestinal (Henthorn et. al., J.Biol. Chem., 263, 12020-12027, 1988). The sequence and complexity of the AP genes from other vertebrates and lower species are now being elucidated. The biological function of AP isozymes is still unknown. In vitro, the enzymes behave as phosphotransferases at neutral pH. The use of phosphate acceptor molecules (diethanolamine, tris, 2-amino-2-methyl-1- propanol) in the buffered substrate solutions increases the reaction rates and, thus, the sensitivity of assays based on AP determinations. ALP is commonly used as a label in immunoassays such as ELISA, and in blotting and histochemistry. Once conjugated to antibodies, antigens, or streptavidin, its low backgrounds and linear reaction rate enables increased sensitivity over extended incubation times. It can be used with a variety of substrates producing precipitated or soluble chromogens, or with chemiluminescent substrates for enhanced sensitivity.

Source:
Alkaline phosphatase, from Calf Intestine


Purity:
≥95%

Activity:
~2000u/mg protein

Form:
Supplied as a lyophilized powder. No preservative added. Reconstitute with sterile ddH2O or diute buffer.

Unit Definitions:
Glycine Units:
The amount of enzyme causing the hydrolysis of one micromole of p-Nitrophenyl phosphate/minute at pH 9.6 and 25°C (glycine buffer).
DEA Units:
The amount of enzyme causing the hydrolysis of one micromole of p-Nitrophenyl phosphate/minute at pH 9.8 and 37°C (diethanolamine buffer).
Unit Conversion:
One Glycine unit as described above is equivalent to ~three DEA units at pH 9.8 and 37°C.

Storage and Stability:
Lyophilized and reconstituted products are stable for 6 months after receipt at -20°C. Reconstitute with sterile ddH2O or diute buffer. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.