Description
Key facts
Sample types:Cell culture extracts, Cell culture media, Cell culture supernatant,
Assay type:Enzyme activity,
Range:5000 - 10000 Cells/well,
Assay Platform:Microscope
Product details:
The Alkaline Phosphatase (AP) Staining Kit (ab284936) (K2035) provides a quick and easy method to stain Alkaline Phosphatase in pluripotent stem cells. It is based on the ability of Alkaline Phosphatase to hydrolyze the phosphate group on the substrate, 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) to form a blue colored intermediate. Nitro Blue Tetrazolium (NBT) then oxidizes the intermediate to form an insoluble, dark blue dimer. Because stem cells/pluripotent stem cells have increased AP activity, the AP Staining Kit can be used to identify and screen Alkaline Phosphatase activity in these cells.
The staining method is simple, rapid, cost-effective and reliable. The entire procedure can be performed in the same microtiter plate and does not require any extra steps such as fixing or harvesting the cells. The assay can be also used as the initial step to test the pluripotency of cells and to monitor osteogenic differentiation of pluripotent/stem cells.
This product is manufactured by BioVision, an Abcam company and was previously called K2035 Alkaline Phosphatase Staining Kit. K2035-50 is the same size as the 50 test size of ab284936.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Alkaline phosphatase also known as ALP is an enzyme with significant biological roles. It mechanically functions by catalyzing the hydrolysis of phosphate groups from various molecules including nucleotides proteins and alkaloids under alkaline conditions. ALP is a metalloenzyme possessing zinc and magnesium ions at its active site to aid in the reaction. This enzyme has a molecular mass of approximately 86 kDa and exists in dimeric form. ALP is expressed in many tissues with high levels in liver bone kidney and the placenta.
Biological function summary
ALP plays an important role in maintaining mineral balance and bone metabolism by breaking down pyrophosphate an inhibitor of mineralization. It facilitates the regulation of phosphate metabolism by contributing to phosphorus and calcium homeostasis. ALP is not part of a larger complex but interacts with calcium and phosphate ions to aid in the formation and resorption of bone mineral tissue. It serves as a marker in bone growth and liver function due to its prominent activity in those tissues visible in assays like the ALP staining method used to gauge enzyme presence and action.
Pathways
Alkaline phosphatase activity plays an essential role in the metabolic pathways related to bone mineralization and liver function. In bone tissue the enzyme participates in the calcification process important for bone strength and development by regulating inorganic phosphate availability. The liver expression of ALP is involved in bile excretion pathways where it dephosphorylates bile acids aiding in their conversion and recycling. It has a significant relationship with proteins such as osteocalcin in the bone mineralization pathway and bile acid transporters in the liver.
Alkaline phosphatase levels serve as diagnostic indicators for bone and liver diseases. High levels of ALP are often associated with bone disorders like Paget's disease and liver conditions like cholestasis. In these disorders ALP activity rises due to increased enzyme synthesis by osteoblasts in bone diseases or impaired bile flow in liver diseases. Furthermore the liver enzyme gamma-glutamyl transferase (GGT) is often measured alongside ALP to differentiate the cause of elevated ALP levels particularly to assess liver pathology.
