Description
Key facts
Detection method:Colorimetric
Product details:
Alkaline Phosphatase Staining Kit (Purple) (ab242286) uses a convenient method to identify and monitor stem cells by staining for alkaline phosphatase activity.
Pluripotent stem cells typically have higher levels of alkaline phosphatase activity than differentiated cell types (although osteoblasts retain this activity). Alkaline phosphatase staining / activity is often used as a biomarker of stem cells.
In this alkaline phosphatase staining kit, alkaline phosphatase acts on a substrate to form a purple colored chemical, producing a purple color in stem cells that is imaged with light microscopy.
The kit can be used with any tissue section or cell type that have alkaline phosphatase activity. It is not compatible with paraffin-embedded sections.
The alkaline phosphatase staining protocol is simple:
- prepare samples (the kit protocol is written for use with ES cells; the kit can be used with any cell or tissues with alkaline phosphatase activity)
- add fixing solution to cells or tissues, incubate for 2 mins, and wash with PBST
- add staining solution and incubate for 15-30 mins
- wash with PBS and image
Alkaline Phosphatase Staining Kit (Red)
ab242286
is an alternative to this kit, using an equivalent method, with a different colored dye. Generally, the red kit gives cleaner staining.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°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.
