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The protocol and study design were approved by the Executive Committee of the Cheyenne River Sioux Tribe Tribal Council (Tribal Resolution number: E-302-08-CR and extended under E-343-2009-CR) and by the University of New Mexico Health Sciences Center Human Research Protection Office (HRPO number: 08-486).
Hydrogen production assay: Hydrogen peroxide production of neutrophil was assessed based on the horseradish peroxidase (HRPO) mediated oxidation of phenol red that resulted in formation of a compound demonstrating increase absorbance at 610 nm (Pick and Keisari, 1980).
Labile Po (LPo) was extracted with 0.5 M NaHC[O.sub.3] (pH 8.5), moderately labile Po (MLPo) was extracted with 1M [H.sub.2]S[O.sub.4] and 0.5M NaOH, moderately resistant Po (MRPo) was dissolved in 0.5M NaOH without any precipitation under pH 1.8, and highly resistant Po (HRPo) was dissolved in 0.5M NaOH with precipitation under pH 1.8:
It uses Hot Rolled Pickled Oiled Coils (HRPO) bought from other companies to be processed into CRC.
Solution 4- 20 mg horseradish peroxidase (HRPO; Sigma P-8375) in 0.5 ml deionized and distilled water
The blot was washed thrice in blotting buffer and reacted with horseradish peroxidase (HRPO) conjugated rabbit anti mouse IgG (Sigma; 1:1000 in blocking buffer) for 1 h.
DTT = dithiothreitol, H&E = haematoxylin and eosin, PAS = periodic acid Schiff, HID = high iron diamine, CsCl = caesium chloride, GuHCl = guanidinium chloride, EDTA = ethylenediaminetetra-acetic acid disodium salt, NEM = N-ethylmaleimide, PMSF = phenylmethysulphonyl fluoride, SDS = sodium dodecyl sulphate, PAGE = polyacrylamide gel electrophoresis, NaCl = sodium chloride, HRPO = horseradish peroxidase, TBST = tris-buffered saline tween-20, TBS = tris-buffed saline tween, [V.sub.i] = included volume, [V.sub.o] = void volume.
The macroarrays were subjected to stringent wash conditions (three 30-min washes with 2x SSC containing 1 g/L SDS at 50[degrees]C; one 30-min wash with 0.2x SSC containing 1 g/ L SDS at 45[degrees]C; and one 1-h wash in 2x SSC at room temperature) and developed with DNA Detector HRPO reagents (KPL) essentially according to the manufacturer's instructions.
The assay was based on horseradish peroxidase (HRPO)mediated oxidation of phenol red by [H.sub.2][O.sub.2], leading to the formation of a compound that absorbs at 610 nm.
Posteriormente se agregaron 50 [micron]l de solucion conjugado de antigeno p26 con peroxidasa de rabano picante (HRPO), se mezclaron y se incubaron por 30 minutos a 37[grados]C, tras tres lavados para eliminar los restos de solucion se colocaron 100 pl de solucion sustrato, se incubaron por 15 minutos a temperatura ambiente.
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- HRR test