Alkaline Phosphatase Assay Kit (Fluorometric) (ab83371) is a
highly sensitive kit to detect alkaline phosphatase (ALP) activity in serum and
other biological samples. In the alkaline phosphatase assay protocol, ALP
cleaves the phosphate group of the non-fluorescent 4-Methylumbelliferyl
phosphate disodium salt (MUP) substrate, resulting in an increased fluorescent
signal (Ex/Em = 360nm/440nm) when the substrate is dephosphorylated.
The kit is an ultra sensitive, simple, direct and HTS-ready
assay designed to measure ALP activity with a detection sensitivity ~1 μU, more
sensitive than colorimetric assays.
Alkaline phosphatase assay protocol summary:
- add samples and standards to wells
- add reaction mix to wells
- add ALP enzyme solution to standard wells (not to samples)
- incubate for 30 min at room temp
- add stop solution
- analyze with microplate reader
Technical Note: ALP is unlikely to be detectable in healthy
urine but may be elevated in the presence of infection.
Related alkaline phosphatase assay products
For a colorimetric alkaline phosphatase assay kit, we recommend Alkaline
Phosphatase Assay Kit (Colorimetric)ab83369. which is based on the
p-nitrophenyl phosphate (pNPP) assay substrate. pNPP turns yellow (405 nm) when
dephosphorylated by ALP as part of an ALP activity assay.
For a luminescent alkaline phosphatase assay, we
recommend Alkaline
Phosphatase Assay Kit (Luminometric)ab233466, which is based on the ability
of alkaline phosphatase to remove the phosphate group from D-luciferin
phosphate. This releases the luciferin to be processed by luciferase.
For alkaline phosphatase staining, we recommend Alkaline
Phosphatase Staining Kit Alkaline
Phosphatase Staining Kitab284936, which is based on the substrate,
5-Bromo-4-chloro-3-indolyl phosphate (BCIP) substrate. BCIP is processed by
alkaline phosphatase to form a blue colored intermediate. Nitro Blue
Tetrazolium (NBT) then oxidizes the intermediate to form an insoluble, dark
blue dimer, which can be viewed by microscopy.
This product is manufactured by BioVision, an Abcam company
and was previously called K422 Alkaline Phosphatase Activity Fluorometric Assay
Kit. K422-500 is the same size as the 500 test size of ab83371.
Alkaline phosphatase (ALP) catalyzes the hydrolysis of
phosphate esters in alkaline buffer and produces an organic radical and
inorganic phosphate. The change in alkaline phosphatase level and activity is
associated with many diseases in the liver and bones.
The Safety Datasheet for this product has been updated for
certain countries. Please check the current version in the Support and
downloads section.
