To measure neutrophil myeloperoxidase (MPO) activity, a portion of the resected cecum was freeze-dried and homogenized in 1 mL of 50 mM pH 6.0 potassium phosphate buffer containing 0.5% hexadecyl trimethyl ammonium bromide. The tissues were disrupted with both sonication and freeze-thaw cycles, and then centrifuged. The supernatant was aliquoted and its protein concentration was measured (Nanodrop 2000C, ThermoScientific). MPO activity in the supernatants was determined using TMB peroxidase substrate (KPL) followed by measurement of the absorbance of the samples at 450 nm using a plate reader. MPO activity (in units per milligram of total protein) was calculated according to the standard curve generated using purified MPO from human leukocytes (Sigma).
Human leukocytes
Manufactured by Merck Group
Human leukocytes are a type of white blood cell that play a crucial role in the immune system. They are responsible for defending the body against infections and diseases. Leukocytes can be further classified into different subtypes, each with specific functions in the immune response.
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Lab products found in correlation
2 protocols using human leukocytes
1
Quantifying Neutrophil Myeloperoxidase Activity
2
Quantifying Intestinal Myeloperoxidase Activity
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To measure neutrophil myeloperoxidase (MPO) activity, a portion of the resected ileum was freeze-dried and homogenized in 1 mL of 50 mmol/L PBS containing 0.5% hexadecyl trimethyl ammonium bromide and 5 mmol/L EDTA. The tissues from each mouse were disrupted with both sonication and freeze-thaw cycles. MPO activity in the centrifuged supernatant was determined using tetramethylbenzidine (TMB) peroxidase substrate (KPL, Gaithersburg, MD) followed by measuring the absorbance of the samples at 450 nm using a plate reader. MPO activity units were calculated according to the standard curve generated using purified MPO from human leukocytes (Sigma, St. Louis, MO).
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