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Alkaline Phosphatase Staining Kit (Red) (ab242286) uses a convenient method to identify and monitor stem cells by staining for alkaline phosphatase activity.

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Immunocytochemistry - Alkaline Phosphatase Staining Kit (Red) (AB242286), expandable thumbnail

Publications

  • Cell reports 40:1110382022
    HMCES modulates the transcriptional regulation of nodal/activin and BMP signaling in mESCs.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Tao Liang,Jianbo Bai,Wei Zhou,Hao Lin,Shixin Ma,Xuechen Zhu,Qinghua Tao,Qiaoran Xi
    PubMed 35830803
  • Experimental biology and medicine (Maywood, N.J.) 247:1379-13872022
    Mouse embryonic stem cells require multiple amino acids.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Boyang Zhang,Jian Feng
    PubMed 35611795

Key facts

Detection method

Colorimetric

Associated Products

Select an associated product type

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Alternative names

What's included?

100 Test
Components
AP Staining Solution A
1 x 20 mL
AP Staining Solution B
1 x 20 mL
Fixing Solution
1 x 50 mL

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Alkaline Phosphatase Staining Kit (Red) (ab242286) uses a convenient method to identify and monitor stem cells by staining for alkaline phosphatase activity.

Alternative names

Key facts

Detection method

Colorimetric

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Alkaline Phosphatase Staining Kit (Red) (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 red colored chemical, producing a red 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 (Purple) Alkaline Phosphatase Staining Kit (Purple) ab242287 is an alternative to this kit, using an equivalent method, with a different colored dye. Generally, this kit gives cleaner staining.

Supplementary info

Activity summary

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.

Associated diseases and disorders

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.

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1 product image

  • Immunocytochemistry - Alkaline Phosphatase Staining Kit (Red) (ab242286), expandable thumbnail

    Immunocytochemistry - Alkaline Phosphatase Staining Kit (Red) (ab242286)

    AP staining of ES Cells.

    Murine embryonic stem cells (ES-D3) are maintained in an undifferentiated stage on gelatin-coated dishes in the presence of LIF, as indicated by the high AP activity. To induce differentiation, LIF was withdrawn over a period of several days; various differentiation events were observed (cells became flattened and enlarged with reduced proliferation). At the end of day 5, AP staining of undifferentiated cells was performed as described in the Assay Protocol.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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