Alkaline phosphatase, Human (HEK293)  (Synonyms: Apase, Human (HEK293))

Alkaline phosphatase (Apase), Human (HEK293) is an orally active membrane-bound glycoprotein that catalyzes the hydrolysis of phosphate monoesters at alkaline pH. Alkaline phosphatase reduces myeloperoxidase activity and bacterial translocation. Alkaline phosphatase improves survival rate of mice infected with E. coli. Alkaline phosphatase improves TNBS-induced colon inflammation^{[1][2][3][4][5][6]}.

Instructions
1. Dissolve the enzyme in 30 U/μL 10 mM Tris-HCl (pH8.0), 1 mM MgCl₂, 50 mM KCl, 0.1 mM ZnCl₂. Fresh preparation is recommended.
2. Add the following reagents to a 1.5 mL centrifuge tube to dephosphorylate DNA:
1) 1-20 pmol DNA fragment
2) 1-2 μL Alkaline Phosphatase
3) 5 μL 500 mM Tris-HCl pH 9.0, 10 mM MgCl2
4) Add sterile ddH₂O to 50 μL
Note: Single-stranded DNA or DNA with 5' protruding ends is recommended to be incubated at low temperature, while blunt-ended DNA or DNA with 3' protruding ends is recommended to be incubated at higher temperature.
3. Mix well and incubate at 37°C or 50°C for 30 min. Alternatively, incubate at 37°C for 15 min, then incubate at 50°C for 15 min.
4. Extract twice with phenol/chloroform/isoamyl alcohol (25:24:1).
Note: Before phenol extraction, it is recommended to incubate it at 56°C for 30 min in a solution containing 5 mM EDTA, 0.5% SDS, and 50 μg/mL Proteinase K to completely inactivate alkaline phosphatase. Incubate at 75°C for 10 min before phenol extraction for better inactivation. 5. Extract with chloroform/isoamyl alcohol (24:1).
6. Add 2.5 μL 3 M NaCl (final concentration 150 mM), 125 μL (2.5 times volume) pre-cooled ethanol, mix well, and place at -20°C for 30-60 min to precipitate DNA.
7. Centrifuge, completely remove ethanol, and dissolve DNA in an appropriate amount of deionized water (<20 μL).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Alkaline phosphatase (1.5 units; i.v.) increases the survival rate of mice injected with a lethal dose of E. coli bacteria^[1].
Alkaline phosphatase (0.15 IU/g; i.v.; once; either 5 min before or 15 min after cecal ligation and puncture) reduces cytokine concentrations in plasma and peritoneal lavage fluid, decreases transaminase activity in plasma and myeloperoxidase activity in the lung in a murine cecal ligation and puncture model of polymicrobial sepsis^[2].
Alkaline phosphatase (5000 U; i.p.) significantly reduces postoperative peritoneal adhesion formation in mice^[3].
Alkaline phosphatase (700 U/kg day; p.o. or intrarectal administration) ameliorates TNBS-elicited colonic inflammation and reduces bacterial translocation in rats^[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

9001-78-9

[Alkaline phosphatase, Human (HEK293)]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Beumer C, et al. Calf intestinal alkaline phosphatase, a novel therapeutic drug for lipopolysaccharide (LPS)-mediated diseases, attenuates LPS toxicity in mice and piglets. J Pharmacol Exp Ther. 2003 Nov;307(2):737-44.  [Content Brief]

  [2]. van Veen SQ, et al. Bovine intestinal alkaline phosphatase attenuates the inflammatory response in secondary peritonitis in mice. Infect Immun. 2005 Jul;73(7):4309-14.  [Content Brief]

  [3]. Hyoju SK, et al. Intestinal alkaline phosphatase decreases intraperitoneal adhesion formation. J Surg Res. 2017 Feb;208:84-92.  [Content Brief]

  [4]. Martínez-Moya P, et al. Exogenous alkaline phosphatase treatment complements endogenous enzyme protection in colonic inflammation and reduces bacterial translocation in rats. Pharmacol Res. 2012 Aug;66(2):144-53.  [Content Brief]

  [5]. Ujjawal Sharma, et al. Alkaline Phosphatase: An Overview. Indian J Clin Biochem. 2014 Jul; 29(3): 269-278.  [Content Brief]

  [6]. Chirambo GM, et al. The role of alkaline phosphatase in intracellular lipid accumulation in the human hepatocarcinoma cell line, HepG2. Exp Mol Pathol. 2017 Apr;102(2):224-229.  [Content Brief]