Description
Key facts
Detection method:Luminescent,
Sample types:Suspension cells, Adherent cells,
Assay type:Quantitative,
Assay Platform:Microplate reader
Product details:
Alkaline Phosphatase Assay Kit (ab233466) uses D-luciferin phosphate as the luminogenic phosphatase substrate to quantify alkaline phosphatase activity in solutions and cells.
D-luciferin phosphate is not recognized by luciferase until its phosphate group is removed to give luciferin. This kit provides all the essential components with an optimized assay protocol which is compatible with HTS liquid-handling instruments. The assay can be readily performed in a 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.
The high sensitivity makes the kit ideal for the assays that require low detection limit.
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 or ALKPHOS is an enzyme that removes phosphate groups from various molecules like nucleotides and proteins. This enzymatic activity requires alkaline conditions with a pH optimum around 8–10. ALP is anchored to the cell membrane and exists in several isoforms each with a mass around 57-70 kDa. Different tissues express specific isoforms such as bone liver kidney and placenta. Osteoblasts for example show high levels of alkaline phosphatase expression often detected by techniques like ALP staining or alkaline phosphatase staining.
Biological function summary
Alkaline phosphatase plays important roles in dephosphorylation processes. Particularly in bone it is associated with osteoblast activity where it is important for bone mineralization. The enzyme is not known to be part of a large protein complex but its activity aids in the conversion of inorganic pyrophosphate into phosphate which promotes the deposition of calcium phosphate in bone. ALP staining and ALP staining osteoblast can be used in laboratory settings for studying the mineralization and function of osteoblasts.
Pathways
Alkaline phosphatase is key in the regulation of the phosphate group levels within the organism particularly in the bone mineralization process. It participates in the vitamin D metabolic pathway which influences calcium and phosphorus homeostasis. Its function has a dynamic relationship with proteins like NPP1 which regulates levels of inorganic pyrophosphate acting counter to alkaline phosphatase's phosphate-producing activity.
Defects in alkaline phosphatase activity are linked to conditions like hypophosphatasia and hyperphosphatasia where mineralization processes are disrupted. In hypophosphatasia decreased ALP activity leads to improper bone formation due to accumulation of inorganic pyrophosphate. This interaction is notable alongside proteins such as tissular nonspecific alkaline phosphatase. On the other hand elevated ALP levels often indicate liver or bone disease pathology where increased enzyme levels reflect altered tissue turnover.
