General description

Embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass (ICM) of preimplantation embryos and are capable of unlimited, undifferentiated proliferation in vitro under appropriate cell culture conditions. Undifferentiated embryonic stem cells are characterized by high expression levels of stage-specific embryonic antigens (SSEA-1, 3 and 4), tumor rejection antigens (TRA-1-60 and TRA-1-81), OCT-4 and alkaline phosphatase. These undifferentiated markers are downregulated upon spontaneous or induced differentiation.

Alkaline phosphatase (ALP) is a hydrolase enzyme responsible for dephosphorylating molecules such as nucleotides, proteins, and alkaloids under alkaline conditions. The enzyme is present within all tissues of the body but is elevated in cells of the liver, kidney, bone, placenta, embryo and under specific disease states. Under alkaline conditions (pH>10), ALP can catalyze the hydrolysis of p-nitrophenylphosphate (p-NPP) into phosphate and p-nitrophenol, a yellow colored by-product of the catalytic reaction. The amount of p-nitrophenol produced is proportional to the amount of alkaline phosphatase present within the reaction. The amount of ALP can thus be reliably quantified by reading the amount of p-nitrophenol amassed after the catalytic reaction at 405 nm on a spectrophotometer.

Millipore’s Quantitative Alkaline Phosphatase ES Characterization Kit allows researchers to easily quantify the amount of alkaline phosphatase present within their embryonic stem cell cultures using a convenient 96-well colorimetric enzymatic assay. This kit can be used in conjunction with Millipore’s Alkaline Phosphatase Detection kit (Cat. No. SCR004) to fully characterize the levels of alkaline phosphatase present within an embryonic stem cell culture.

Application

Research Category
Stem Cell Research

This Quantitative Alkaline Phosphatase Embryonic Stem (ES) Characterization Kit easily quantifies the amount of alkaline phosphatase present within their embryonic stem cell cultures using a convenient 96-well colorimetric enzymatic assay.

Components

p-NPP Substrate Concentrate (50X): (Part No. ES008-200UL) 200 μL

p-NPP Buffer: (Part No. ES011-10ML) 10 mL

Reaction Stop Solution: (Part No.CS200653) 5 mL

1X Wash Solution: (Part No. CS200652) 2 X 125 mL

Recombinant Alkaline Phosphatase Standard: (Part No. CS200654) 0.5 μg (10 μg/mL)

Storage and Stability

All reagents are to be stored at 2∘ to 8∘C until expiration date. p-NPP substrate concentrate and p-NPP buffer are light sensitive and should not be exposed to light. Reaction stop solution contains 1N sodium hydroxide and is corrosive. Avoid skin contact and inhalation.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.