After collection, the device was centrifuged at 4,000 rpm at 4°C for 5 min to recover clean saliva, the intermediary from the upper reservoir of tampon, and the pellet reservoir. Saliva (1 mL) was added to tubes containing chilled absolute Ethanol (1 mL), vortexed, and stored overnight at -20°C. The precipitated antibodies and globulins were pelleted by centrifugation at 10,000 rpm for 5 min. The pellet was suspended in 400 µL of saline borate buffer Ph 7.2 with 1 mg/mL of BSA (Serum Bovine Albumin, Sigma®), resulting in 2.5× concentrated saliva, which was then stored at -20°C until use.
Serum bovine albumin
Manufactured by Merck Group
Sourced in United States
Serum bovine albumin is a laboratory reagent derived from bovine serum. It is a protein that is commonly used in cell culture applications to support cell growth and maintenance.
Automatically generated - may contain errors
Lab products found in correlation
Beta amylase, by Merck Group (2 mentions)
Sybr greener qpcr supermix universal kit, by Thermo Fisher Scientific (1 mentions)
Bradford reagent, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
Mgcl2, by Merck Group (1 mentions)
Genistein, by Merck Group (1 mentions)
Goat anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Enhanced chemiluminescent ecl kit, by GE Healthcare (1 mentions)
Ponceau s stain, by Merck Group (1 mentions)
Z36hk, by Hermle (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Malondialdehyde mda, by Cayman Chemical (1 mentions)
7 protocols using serum bovine albumin
1
Salivary Antibody Concentration by Ethanol Precipitation
2
Bradford Assay for Protein Quantification
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Total protein concentration in lung homogenates was determined by Bradford colorimetric assay at 595 nm [62 (link)] employing serum bovine albumin (Sigma-Aldrich, US) (0–1.4 mg/mL) as standard.
3
In Vitro Hepatoprotective Assay
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Genistein (5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one, CAS number 446-72-0), serum bovine albumin, HEPES, EDTA, EGTA, DTT, PMSF, protease inhibitor cocktail, Ponceau S stain, and Bradford reagent were procured from Sigma, St. Louis, USA. DMSO, KCl, MgCl2, and H2O2 were purchased from Merck, India. ALT, AST, and ALP estimation kits were purchased from Beckman Coulter, Ireland. TRIzol, First-Strand Synthesis Supermix for qRT-PCR and SYBR GreenER qPCR Supermix Universal kit were purchased from Invitrogen, California, USA. Primary antibodies such as rabbit polyclonal IgG SOD1, actin, and secondary antibody goat anti-rabbit IgG-HRP were procured from Santa Cruz Biotechnology, California. Purified mouse anti-Hsp70 antibody was purchased from BD Biosciences, USA. Enhanced chemiluminescent (ECL) kit was procured from GE Healthcare, UK.
4
Biochemical Assays for Muscle Tissue
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For biochemical assays, muscle tissue (100 mg/mL) was homogenized in RIPA buffer (Sigma-Aldrich, Saint Louis, MO, USA) and centrifuged at 1600× g for 10 min at 4 °C (Z 36 HK, Hermle-Labortechnik, Wehingen, Germany). Supernate was collected for determining the thiobarbituric-acid-reactive substances (TBARS), total peroxidase, and glutathione peroxidase activities.
TBARS levels were calculated based on the standard curve of malondialdehyde (MDA) (Cayman Chemical, Ann Arbor, MI, USA) (0 to 50 µM) and followed the protocol described previously [11 (link)]. TBARS levels (μM) in muscle tissue were normalized by protein concentration (mg/mL) and expressed as μM/mg protein.
Total peroxidase activity was measured by the disappearance of hydrogen peroxide (H2O2) at 240 nm [12 (link)], whilst the total glutathione peroxidase (GPx) activity was determined according to previously standardized methods [13 (link)]. Total peroxidase and GPx activities were normalized by protein concentration (mg/mL) and expressed in mU/mg protein.
Total protein concentration of the muscle homogenates was determined by Bradford colorimetric assay at 595 nm (Sigma-Aldrich, Saint Louis, MO, USA) employing serum bovine albumin (Sigma-Aldrich, Saint Louis, MO, USA) (0 to 1.4 mg/mL) as standard [14 (link)].
TBARS levels were calculated based on the standard curve of malondialdehyde (MDA) (Cayman Chemical, Ann Arbor, MI, USA) (0 to 50 µM) and followed the protocol described previously [11 (link)]. TBARS levels (μM) in muscle tissue were normalized by protein concentration (mg/mL) and expressed as μM/mg protein.
Total peroxidase activity was measured by the disappearance of hydrogen peroxide (H2O2) at 240 nm [12 (link)], whilst the total glutathione peroxidase (GPx) activity was determined according to previously standardized methods [13 (link)]. Total peroxidase and GPx activities were normalized by protein concentration (mg/mL) and expressed in mU/mg protein.
Total protein concentration of the muscle homogenates was determined by Bradford colorimetric assay at 595 nm (Sigma-Aldrich, Saint Louis, MO, USA) employing serum bovine albumin (Sigma-Aldrich, Saint Louis, MO, USA) (0 to 1.4 mg/mL) as standard [14 (link)].
5
Cellulose Hydrolysis Enzyme Assay
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The reagents p-nitrophenyl β-d -glucopyranoside (ρNPβGlu), methylumbelliferyl-β-glucopyranoside (MUβGlu), octyl-β-glucopyranoside (octylβGlu), gentiobiose, laminaribiose, cellobiose, cellotriose, cellotetraose, cellopentaose, microcrystalline cellulose (Avicel®, Cat 11365), low viscosity carboxymethylcellulose (CMC, Cat C5678), ρ-nitrophenyl-β-d -galactopyranoside (ρNPβgal), methylumbeliferil-β-d -mannopyranoside (MUβMan), ρ-nitrophenol, 4-methylumbelliferone, cellulases from T. reesei ATCC 26921 (Cat. C8546) and the molecular markers Blue dextran (Cat. D4772), Cytochrome C (Cat. 12 kDa), carbonic anhydrase (Cat. C7025), serum albumin bovine (Cat. A8531), alcohol dehydrogenase (Cat. A8656), and beta amylase (Cat. A8781) were purchased from Sigma-Aldrich Company (St. Louis, Missouri, USA). Trypsin (Cat. V511A) for mass spectrometry was acquired from Promega Corporation (Madison, Wisconsin, USA). Glucose oxidase-based glucose quantitation reagent was acquired from Bioclin (Minas Gerais, Brazil). Other reagents used in this work were analytical grade.
6
Praziquantel Effectiveness on Liver Enzymes
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Praziquantel was supplied by Merck S.A., Mexico, Mexico. Total bilirubin and alkaline phosphatase kits (Inmesco kits) were purchased from Inmesco, Wied, Germany. Transaminases kits were procured from Quibasa Quimica Basica Ldta, Belo Horizonte, Brasil. The following were products of Sigma-Aldrich, St Louis, USA: Ellman's reagent, serum albumin bovine (SAB), trichloroacetic acid (TCA), thiobarbituric acid (TBA), chemicals used for the preparation of the Biuret reagent and buffers. Methanol and others chemicals were of analytical grade. For parasitological study, we used a stereo-microscope (Zeiss, Hamburg, Germany) and a light microscope (Leica, Wetzlar, Germany). For biochemical analyses, we used the Genesys 20 spectrophotometer (Sigma-Aldrich, St Louis, USA).
7
SEC Analysis of EPS Samples
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SEC was performed using (i) a liquid chromatograph with pump (LC-20AD), (ii) a fluorescence detector (RF-20AXS), (iii) an auto sampler (SIL-20AHT), and (iv) a communication module (CMB-20A). The molecular weights (MW) of the EPS samples were estimated by passing 15 μL of the filtered samples through an analytical scale SEC column (OHpak SB-804 HQ) following its guard column (OHpak SB-G). Tris buffer (25 mM Tris, pH 7.0 ± 0.1) was used as the mobile phase. The SEC column was calibrated using transferrin, serum albumin bovine, myoglobin and beta amylase obtained from Sigma Aldrich (the details are provided in Supporting Information (SI)).
not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
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