Product Introduction
Alkaline phosphatase (AP/ALP/AKP/ALKP/ALPase/Alk Phos), also known as alkaline phosphatase (EC 3.1.3.1), can catalyze the hydrolysis of phosphate bonds under alkaline conditions. In mammals, the activity of alkaline phosphatase in the liver, bile duct, kidney, bone and placenta is relatively high. Common alkaline phosphatases include intestinal alkaline phosphatase (ALPI), non-tissue-specific alkaline phosphatase (ALPL) and placental alkaline phosphatase (PLAP). Except for placental alkaline phosphatase (placental isoform), other endogenous alkaline phosphatases are easily inactivated after heating. The common Calf Intestinal Alkaline Phosphatase (CIAP/CIP) is widely used for labeling secondary antibodies and ultimately for the detection of proteins and nucleic acids. It is also commonly used for dephosphorylation of the 5´ and 3´ ends of DNA or RNA, especially dephosphorylation of the 5´ end of plasmids to avoid plasmid self-ligation.
In stem cells, such as iPS, alkaline phosphatase activity is very high and is often used as a marker of successful iPS induction. In addition, the activity of alkaline phosphatase in differentiated colon cancer cells will also increase significantly, which is used as a qualitative and quantitative indicator of the degree of differentiation of colon cancer cells. In addition, the increase in serum alkaline phosphatase, called hyperalkaline phosphatasemia, is considered to be related to malignant biliary obstruction, primary biliary cirrhosis, Hepatobiliary diseases such as primary sclerosing cholangitis, hepatic lymphoma and hepatic sarcoidosis are closely related. Elevated serum alkaline phosphatase activity is also closely related to bone formation, because alkaline phosphatase is a by-product of osteoblasts. Low serum alkaline phosphatase activity is also associated with some diseases. The alkaline phosphatase activity in the serum of children and pregnant women is higher than that of ordinary people. The alkaline phosphatase activity in serum ranges from 20-140U/L.
Principle
Para-nitrophenyl phosphate (pNPP) is a commonly used chromogenic substrate for phosphatase. Under alkaline conditions, it can generate para-nitrophenol (pNP) under the action of alkaline phosphatase. p- nitrophenol is a yellow product under alkaline conditions, and the absorbance can be detected at 400-415nm. The darker the yellow color of the product, the higher the activity of alkaline phosphatase, and vice versa. Based on this, the activity level of alkaline phosphatase can be calculated by colorimetric analysis.
Components
Serial number | Components | Size(100T) | Storage |
Reagent 1 | Assay Buffer | 15ml | -20℃ |
Reagent 2 | Chromogenic Substrate | 2 tubes | -20℃, keep away from light. |
Reagent 3 | p -nitrophenol solution (10mM) | 0.1ml | -20℃, keep away from light. |
Reagent 4 | Reaction stop solution | 12ml | -20℃ |
Consumable 1 | Microplate(96 wells) | 1 plate | RT |
Consumable 2 | Plate Sealer | 2 pieces | RT |
Storage
The unopened kit can be stored at -20℃ for 12 months.
Notes
This product is intended for scientific research use only by professionals and must not be used for clinical diagnosis or treatment, in food or drugs, or stored in ordinary residences.
