We used the catalase assay developed by Iwase et al.16 (link) with slight modifications. Bacterial strains were grown overnight in 1% soytone at 37°C with shaking. Optical density at 600 nm (OD600) was recorded to estimate bacterial concentration and 2 mL of culture (6–8 × 109 bacteria) was centrifuged; the supernatant was removed; and the cell pellet was resuspended in 100 μL of normal saline (9 g/L sodium chloride; Quality Biological, Gaithersburg, MD) and transferred into 13 × 100 mm borosilicate glass tubes. One-hundred microliters of 1% Triton X-100 and 100 μL of 35% stabilized hydrogen peroxide were added. The tubes were left untouched at room temperature until the foaming stopped (about 2 minutes). The height of the foam in the tube was then recorded. A standard curve was also created using dilutions of commercially available bovine catalase (Sigma-Aldrich; 3,000 units/mg), and the final results were expressed as units of catalase (U) per 1010 bacterial cells, assuming an OD600 of 1.25 corresponds to 109 bacteria/mL of culture for all strains tested.
Bovine catalase
Manufactured by Merck Group
Sourced in Germany, United States
Bovine catalase is an enzyme isolated from bovine liver. It functions as a catalyst, accelerating the breakdown of hydrogen peroxide into water and oxygen.
Automatically generated - may contain errors
Lab products found in correlation
Epinephrine, by Merck Group (10 mentions)
2 thiobarbituric acid, by Merck Group (6 mentions)
Trichloroacetic acid, by Merck Group (6 mentions)
Sodium chloride (nacl), by Merck Group (4 mentions)
Methanol, by Merck Group (4 mentions)
Cytochrome c, by Merck Group (4 mentions)
Nadph, by Merck Group (4 mentions)
Sodium hydroxide (naoh), by Merck Group (3 mentions)
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Hydrochloric acid (hcl), by Merck Group (3 mentions)
Butylated hydroxyl toluene, by Merck Group (3 mentions)
Hematoxylin, by Merck Group (2 mentions)
Acetic acid, by Merck Group (2 mentions)
Butylated hydroxytoluene, by Merck Group (2 mentions)
2 thiobarbituric acid tba, by Merck Group (2 mentions)
Aspergillus niger glucose oxidase, by Merck Group (2 mentions)
Ethanol, by Merck Group (2 mentions)
Rutin, by Merck Group (2 mentions)
Folin ciocalteu, by Merck Group (2 mentions)
Horseradish peroxidase hrp, by Merck Group (2 mentions)
45 protocols using bovine catalase
1
Catalase Assay for Bacterial Quantification
2
Assessing Oxidative Stress in Cancer Cells
3
Histochemical Analysis Protocol
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Acetic acid, NaCl, Hematoxylin, Eosine, Formaldehyde, Epeniphrine, Bovine catalase, Butulated hydroxytoluene (BHT), trichloroAcetic acid (TCA), 2-Thio-barbituric acid (TBA) NaOH and methanol, were from sigma chemicals Co (Germany). All other chemicals employed were of analytical grade.
4
Calcineurin Activation Assay Protocol
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All chemicals were of analytical reagent
(AR) grade. Ethylene glycol-bis(2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), dithiothreitol (DTT), iodoacetamide,
bovine catalase, tris(hydroxymethyl)aminomethane hydrochloride
(Tris·HCl), tris(hydroxymethyl)aminomethane (Tris·base),
and HEPES were purchased from Sigma-Aldrich (St. Louis, MO, USA).
KCl, MgCl2, CaCl2, and H2O2 (30% solution) were purchased from Columbus Chemical Industries
(Columbus, WI, USA). Dimethylformamide (DMF) was from Jade Scientific
(Canton, MI, USA). Glacial acetic acid and water (HPLC grade) were
from Mallinckrodt Chemicals (Phillipsburg, NJ, USA). Acetonitrile
(HPLC grade) and iodomethane were purchased from EMD Chemicals (San
Diego, CA, USA). Sodium hydroxide (NaOH) and formic acid were from
Spectrum Chemical Mfg. (Gardena, CA, USA). Trifluoroacetic acid (TFA)
was purchased from Pierce (Rockford, IL, USA). Mass spectrometry grade
Glu-C was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Sequencing grade modified trypsin was from Promega (Madison, WI, USA).
Sequencing grade Lys-C was obtained from Roche Diagnostics (Indianapolis,
IN, USA). Human calcineurin was prepared as previously described.27 (link) DMBNHS was prepared via a three-step process
as described previously.31 (link)
(AR) grade. Ethylene glycol-bis(2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), dithiothreitol (DTT), iodoacetamide,
bovine catalase, tris(hydroxymethyl)aminomethane hydrochloride
(Tris·HCl), tris(hydroxymethyl)aminomethane (Tris·base),
and HEPES were purchased from Sigma-Aldrich (St. Louis, MO, USA).
KCl, MgCl2, CaCl2, and H2O2 (30% solution) were purchased from Columbus Chemical Industries
(Columbus, WI, USA). Dimethylformamide (DMF) was from Jade Scientific
(Canton, MI, USA). Glacial acetic acid and water (HPLC grade) were
from Mallinckrodt Chemicals (Phillipsburg, NJ, USA). Acetonitrile
(HPLC grade) and iodomethane were purchased from EMD Chemicals (San
Diego, CA, USA). Sodium hydroxide (NaOH) and formic acid were from
Spectrum Chemical Mfg. (Gardena, CA, USA). Trifluoroacetic acid (TFA)
was purchased from Pierce (Rockford, IL, USA). Mass spectrometry grade
Glu-C was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Sequencing grade modified trypsin was from Promega (Madison, WI, USA).
Sequencing grade Lys-C was obtained from Roche Diagnostics (Indianapolis,
IN, USA). Human calcineurin was prepared as previously described.27 (link) DMBNHS was prepared via a three-step process
as described previously.31 (link)
5
Synthesis and Reactivity of Arylsulfonyl Azides
6
Catalase Activity in Fibroblasts
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Fibroblasts were lysed in PBS by repeated freezing and thawing, in the presence of protease inhibitors. Cat activity was determined by spectrophotometric monitoring the rate of disappearance of H2O2 at 240 nm [34] . A standard curve was obtained with bovine catalase (Sigma-Aldrich, Srl Milan, Italy). Units were normalized for protein content. Results of experiments performed in each donor (n = 3) in triplicate are given as% of relative units of Cat per mg protein ± S.D.
7
Preparation of Bacterial Antimicrobial Supernatant
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Overnight cultures of the potential probiotic isolates cultivated on broth medium were diluted with fresh medium (inoculum size 1% v/v) at 32 °C for 18 h, and the concentration was adjusted to an optical density of 1.6 at 600 nm (~1 × 108 cells/mL). MRS broth (Oxoid, Basingstoke, UK) was used for the cultivation of lactobacilli, and M17 broth (Oxoid, Basingstoke, UK) was used for the cultivation of streptococci. Cultures were centrifuged at 10,000× g for 15 min at 4 °C and the resulting supernatant was designated as a crude cell-free culture supernatant (CCFCS). To neutralize hydrogen peroxide, 1 mg/mL of bovine catalase (Sigma-Aldrich, St. Louis, MO, USA) was added. The pH of each CCFCS was adjusted to 6.5 with 1 mol/L NaOH (Sigma-Aldrich, St. Louis, MO, USA) [21 (link)]. The treated supernatant was designated a BLIS. The 10-fold concentration of the BLIS was obtained using a vacuum rotary evaporator at 40 °C, filtered through a sterile 0.2 μm syringe filter, and stored at −20 °C until further use.
8
Plasma Treatment of hsFB Cells
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Prior to plasma treatment in PBS (500–1000 µL), the hsFB were carefully washed (PBS, 500 µL), and the culture plate was placed under the μAPPJ. The distance between the electrode end and the culture plate well’s bottom in all experiments was kept at 7 mm (Figure 2 A) for direct treatments. For indirect treatments, the buffer or media was separately treated from the hsFB and immediately transferred to the hsFB-containing cell culture plate. The directly and indirectly treated hsFB were incubated as indicated (0–5 min) before buffer was removed and fresh media was added. In addition, the hsFB were incubated with a buffer containing bovine catalase (1000 U/mL, Sigma Aldrich, St. Louis, MO, USA).
9
SDS-PAGE Gel Electrophoresis Reagents
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Acrylamide, methylene-bis-Acrylamide, N,N,N′,N′-tetramethylethylenediamine, and ammonium persulfate for sodium dodecyl sulfate polyAcrylamide gel electrophoresis (SDS-PAGE) were purchased from Bio-Rad. Blue dextran, thyroglobulin, bovine catalase, bovine serum albumin, and lysozyme were purchased from Sigma. HiPrep DEAE FF 16/10 column, Superdex 200 10/300 GL column, and Resource Q column were purchased from General Electrics. All other chemicals were of the best purity available.
10
Malathion Purity Evaluation Protocol
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Malathion (Fyfanon EC 500) was obtained from INNOVA Society size in Tunis, Tunisia at The purity of 96%. 2-Thio-barbituric acid (TBA), bovine catalase, Epinephrine, and butylated hydroxytoluene (BHT) were from Sigma aldrich chemicals Co (Germany). All other chemicals used were of analytical grade.
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