Alkaline Phosphatase Assay - Information
Alkaline Phosphatase Assay Kit (Colorimetric) is designed to measure ALP activity directly in biological samples without pretreatment. The improved method utilizes p-nitrophenyl phosphate that is hydrolyzed by ALP into a yellow colored product (maximal absorbance at 405nm). The rate of the reaction is directly proportional to the enzyme activity.
Applications
For quantitative determination of alkaline phosphatase (ALP) activity using stable p-nitrophenol phosphate substrate.
Alkaline Phosphatase Assay - Key Features
- High sensitivity and wide linear range. Use 5 serum or plasma sample. The detection limit is 2 U/L, linear up to 800 U/L.
- Homogeneous and simple procedure. Simple "mix-and-measure" procedure allows reliable quantitation of ALP activity within 5 minutes.
- Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments.
Alkaline Phosphatase Assay - Data Sheet | |
Kit Includes | Assay Buffer: 50 mL, pH 10.5 Mg Acetate: 1.5 mL 0.2 M pNPP Liquid: 600 mL 1 M Calibrator: 10 mL Tartrazine |
Kit Requires | Pipeting devices and accessories (e.g. multi-channel pipettor) |
Method of Detection | OD405nm |
Detection Limit | 2 U/L |
Samples | Serum, plasma etc |
Species | All |
Protocol Length | 4 min |
Size | 250 tests |
Storage | Store pNPP Liquid and Calibrator at -20°C and other components at 4°C. |
Shelf Life | 12 months |
More Details
Alkaline phosphatase(ALP) catalyzes the hydrolysis of phosphate esters in an alkaline environment, resulting in the formation of an organic radical and inorganic phosphate. In mammals, this enzyme is found mainly in the liver and bones. Marked increase in serum ALP levels, a disease known as hyperalkalinephosphatasemia, has been associated with malignant biliary obstruction, primary biliary cirrhosis, primary sclerosing cholangitis, hepatic lymphoma and sarcoidosis. Simple, direct and automation-ready procedures for measuring ALP activity in serum are becoming popular in Research and Drug Discovery.
Aliases for ALPL Gene
- AlkalinePhosphatase, Liver/Bone/Kidney
- AlkalinePhosphataseLiver/Bone/Kidney Isozyme
- EC 3.1.3.1
- AP-TNAP
- TNSALP
- AlkalinePhosphatase, Tissue-Nonspecific Isozyme
Entrez Gene Summary for ALPL Gene
This gene encodes a member of thealkalinephosphatasefamily of proteins. There are at least four distinct but relatedalkalinephosphatases: intestinal, placental, placental-like, and liver/bone/kidney (tissue non-specific). The first three are located together on chromosome 2, while the tissue non-specific form is located on chromosome 1. The product of this gene is a membrane bound glycosylated enzyme that is not expressed in any particular tissue and is, therefore, referred to as the tissue-nonspecific form of the enzyme. Alternative splicing results in multiple transcript variants, at least one of which encodes a preproprotein that is proteolytically processed to generate the mature enzyme. This enzyme may play a role in bone mineralization. Mutations in this gene have been linked to hypophosphatasia, a disorder that is characterized by hypercalcemia and skeletal defects. [provided by RefSeq, Oct 2015]
UniProtKB/Swiss-Prot for ALPL Gene
PPBT_HUMAN,P05186
- This isozyme may play a role in skeletal mineralization
Protein details for ALPL Gene (UniProtKB/Swiss-Prot)
Protein Symbol:P05186-PPBT_HUMAN
Recommended name:Alkalinephosphatase, tissue-nonspecific isozyme
Protein Accession:P05186
Secondary Accessions:A1A4E7, B2RMP8, B7Z387, B7Z4Y6, O75090, Q2TAI7, Q59EJ7, Q5BKZ5, Q5VTG5, Q6NZI8, Q8WU32, Q9UBK0
Protein attributes for ALPL Gene
Size:524 amino acids
Molecular mass:57305 Da
Cofactor:Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
Cofactor:Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
Quaternary structure:Homodimer.
SequenceCaution:Sequence=BAD93051.1; Type=Erroneous initiation; Note=Translation N-terminally shortened.; Evidence={ECO:0000305};
Post-translational modifications for ALPL Gene
- N-glycosylated.P05186-PPBT_HUMAN
- Glycosylation at Asn 140, Asn 230, Asn 271, Asn 303, and Asn 430NX_P05186
- Modification sites at PhosphoSitePlusP05186