The cells cultured in chamber slides were fixed using an iced fixative solution (3.7 % formaldehyde, 3 % sucrose in PBS 1X) for 20 min at RT, permeabilized with ice 0.5 % Triton X-100 in PBS 1X in ice at 4 °C for 20 min, incubated with 3 % hydrogen peroxide in PBS 1X for 8 min and then maintained in a blocking solution (PBS 1X with 3 % albumin from bovine serum-BSA, Sigma, Milan, Italy) for 1 h in at RT, as previously described [36 (link)]. After this time the slides were incubated overnight at 4 °C in a humidified chamber with specific primary antibody VDR receptor (1:50). Then chamber slides were incubated before with diluted biotinylated secondary antibody solution (Dako Italia, Milan, Italy), then with VECTASTAIN® ABC Reagent (Dako Italia, Milan, Italy), and finally in peroxidase substrate solution (Peroxidase/DAB, Dako Italia, Milan, Italy), and counterstained with Mayer’s hematoxylin. The number of positive cells was calculated as described elsewhere [37 (link)]; briefly, 12 different areas (1 mm2) randomly selected from each section were taken, and the number of signals was determined using ImagePro 3 software (NIH, Bethesda, US). The results are expressed as means ± SD (%).
Albumin from bovine serum bsa
Manufactured by Merck Group
Sourced in United States, Germany
Albumin from bovine serum (BSA) is a laboratory reagent derived from the serum of bovine (cattle) origin. It is a protein that is commonly used in various biological and biochemical applications. The core function of BSA is to serve as a stabilizing agent, blocking agent, and protein source in various experimental protocols.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Merck Group (4 mentions)
Fetuin, by Merck Group (3 mentions)
Ham s f 10 medium, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Euroclone (3 mentions)
D glucose, by Merck Group (3 mentions)
Dimethyl sulfoxide (dmso), by Avantor (2 mentions)
Trifluoroacetic acid hplc grade, by Merck Group (2 mentions)
Dulbecco s modified eagle s medium f12 nutrient mixture ham dmem f12 3 1, by Thermo Fisher Scientific (2 mentions)
N succinyl tri alanyl p nitroanilide stana, by Merck Group (2 mentions)
Procollagen type 1 peptide pip elisa kit, by Takara Bio (2 mentions)
Matrix metalloproteinase 1 mmp 1 human elisa kit, by Abcam (2 mentions)
Yokudelna calibration kit, by JEOL (2 mentions)
Antibiotic antimycotic solution, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Bradford reagent, by Merck Group (2 mentions)
Sucrose, by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
26 protocols using albumin from bovine serum bsa
1
Immunohistochemical Analysis of Vitamin D Receptor
2
Immunofluorescence Analysis of Heparanase and RNA Pol II
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HRECs were seeded on glass coverslips precoated with 5 mg/ml of fibronectin (Gibco) and allowed to grow to semiconfluence in a culture dish. The cells were washed with phosphate buffered saline (PBS) three times and fixed in fresh 4% paraformaldehyde (pH 7–8) for 10 min at room temperature. PBS was composed of 2.89 g of Na2HPO4·12H2O, 8 g of NaCl, 0.2 g of KCl, 0.2 g KH2PO4 and 80 ml of ddH2O. Next, the HRECs were permeabilized in 0.1% of Triton X-100 (Sigma, St. Louis, MO) for 5 min and blocked with 1% albumin from bovine serum (BSA; Sigma) in PBS containing 0.1% Tween 20 (blocking solution) for 60 min at room temperature. The cells were then incubated with rabbit anti-human heparanase antibody (1:300 dilution; Abcam, Cambridge, MA) and mouse anti-human RNA Pol II antibody (1:200 dilution; Abcam) overnight at 4 °C for the expression of heparanase and RNA Pol II, followed by incubation with appropriate secondary antibodies conjugated with Alexa Fluor 488 and Alexa Fluor 555 (1:200 dilution; Boster Biologic Technology, Ltd., Wuhan, China) for 2 h at room temperature. The cells were stained with 100 ng/ml of 4',6-diamidino-2-phenylindole (DAPI; Sigma) for 5 min, mounted with an antifading fluorescence medium (Vector Laboratories, Burlingame, CA), and imaged using a laser scanning confocal microscope (Carl Zeiss, Jena, Germany).
3
Synthesis and Characterization of Chitosan-Based Nanocomposites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All salts and solvents, unless otherwise stated, were obtained from Fisher Scientific (UK). Sodium phosphate monobasic monohydrate, low viscosity chitosan from shrimp, glycerol phosphate disodium salt hydrate, tris(hydroxymethyl)amino-methane [(HOCH2)3CNH2], albumin from bovine serum (BSA), albumin from chicken egg white (OVA) and QuantiPro™ Bicinchoninic Acid Assay Kit were obtained from Sigma-Aldrich (UK). l (+)-lactic acid 90 % solution in water was obtained from Acros Organics (USA). Dialysis membranes (size 10, MWCO 12–14 kDa) were obtained from Medicell International Ltd (UK). MWCTs and SWCTs were purchased from Cheap Tubes Inc. (USA). The MWCTs used in the study were 8–15 nm in diameter and 10–50 µm in length. The SWCTs used were 1–4 nm in diameter and 5–30 µm in length. Di-potassium hydrogen orthophosphate anhydrous (K2HPO4) was acquired from British Drug Houses (UK) and troclosene sodium dehydrate from Guest Medical (UK). Water used in all experiments was purified water. NOSC and chitosan modified hydroxyapatite (HACS) were prepared as described previously [12 (link), 21 (link)].
4
Synthesis and Characterization of Stimuli-Responsive Polymer Nanomaterials
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1-Propanethiol (Aladdin, Shanghai, China, 99%), carbon disulfide (Sigma, Metairie, LA, USA, 99%), potassium ferricyanide (K3Fe(CN)6, Aladdin, Shanghai, China, 99%), 4,4′-azobis (4-cyanovaleric acid) (ACVA, Sigma, Metairie, LA, USA, 98%), 2,2′-Dithiodipyridine (Aladdin, Shanghai, China, 98%), 2-Mercaptoethanol (Sigma, Metairie, LA, USA, 99%). N′-dicyclohexylcarbodiimide (DCC, Sigma, Metairie, LA, USA, 99%), 4-(dimethylamino)pyridine (DMAP, Sigma, LA, USA, 99%), Tris(2,2-bipyridine) dichlororuthenium(II) hexahydrate (Aladdin, Shanghai, China, 98%), N-isopropylacrylamide (NIPAAm, Sigma, LA, USA, 98%) were recrystallized twice in hexane and toluene prior to use. 2-ethyl-1-hexanol (Sigma, LA, USA, ≥98%), ALP and 4-nitrophenyl phosphate (pNPP) (Sigma, Metairie, LA, USA), PEG-bis (N-succinimidyl succinate) (Sigma, Metairie, LA, USA), Chloroauricacid (HAuCl4·H2O, Energy Chemical, Shanghai, China, 98%), sodiumhydroxide (NaOH, Sigma, Metairie, LA, USA) were purchased from Guangfu Technology Development Co. Ltd., Tianjin, China). Albumin from bovine serum (BSA, isoelectric point = 4.6) (Sigma, Metairie, LA, USA, ≥98%) were used as received without further purification.
5
Investigating Myotube Irisin Secretion in Myoblasts from DM Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human satellite cells were isolated from biceps brachii muscle biopsies from 3 DM1, 3 DM2 patients and from three subject with no sign of neuromuscular disease used as controls, as previously described (22 (link)). Myoblasts were grown in HAM’s F10 medium (Sigma-Aldrich) supplemented with 15% FBS (Euroclone), 0.5 mg/mL albumin from bovine serum (BSA, Sigma-Aldrich), 0.5 mg/mL fetuin (Sigma-Aldrich), 0.39 µg/mL dexamethasone, 10 ng/mL epidermal growth factor, 0.05 mg/mL insulin, 3 mg/mL glucose, 100 U/mL penicillin, and 100 µg/mL streptomycin (proliferative medium). For this study, cells from DM and control patients were plated at a density of 60,000 cells per 35 mm dishes. When myoblasts reached 80% of confluence, proliferative medium was replaced by differentiative medium (DMEM/High Glucose supplemented with 7% FBS, in presence of 100 U/mL penicillin, 100 µg/mL streptomycin, and 0.01 mg/mL insulin) to allow myoblasts fusion. Myotubes and conditioned media were harvested after 5 days of differentiation (T5) and irisin concentration was determined after 6 h of incubation in serum and fetuin-free conditioned media.
6
Synthesis of Ag@ZnO Nanostructures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Zinc nitrate hexahydrate, hexamethylenetetramine (HMTA), and silver nitrate were purchased from Sigma-Aldrich with a grade of analytical purity for the synthesis of Ag@ZnO nanostructures. 4-Aminothiophenol (4-ATP), rhodamine 6G (R6G), and albumin from bovine serum (BSA) were purchased from Sigma-Aldrich as SERS detecting samples. The precursor solution for ZnO nanorod fabrication was a mixture of zinc nitrate (25 mM) and HMTA (25 mM). The precursor solution for Ag nanoparticle growth was a silver nitrate aqueous solution (20 mM).
7
Fungi Cultivation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pheaotheca triangularis EXF-206, T. salinum EXF-295, and Wallemia ichthyophaga EXF-5676 were obtained from the University of Ljubljana, Biotechnical Faculty, Department of Biology (Ljubljana, Slovenia). Carbon dioxide 2.5 (purity 99.5%) was supplied by Messer MG (Ruše, Slovenia). Peptone from meat, potassium phosphate, potassium dihydrogen phosphate, sodium carbonate, sodium bicarbonate, and acetic acid were purchased from Merck (Darmstadt, Germany). Sodium pyrophosphate decahydrate (≥99.0%), sodium phosphate monobasic (≥99.0%), sodium phosphate dibasic (≥99.0%), albumin from bovine serum (BSA) (≥98.0%), malt extract, agar, d -(+)-glucose, sodium acetate, Sigmacell, glucose assay reagent, Casein, Hammarsten bovine, trichloroacetic acid (TCA), d -(−)-Salicin (≥99.0%), starch azure, and sodium chloride were supplied from Sigma (Schnelldorf, Germany).
8
Synthesis and Characterization of Targeted Theranostic Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All materials were purchased from commercial suppliers and used without further purification, unless otherwise noted. Cy5.5-NHS was purchased from GE Healthcare (Piscataway, NJ, USA). Succinimidyl iodoacetate (SIA), 2-iminothiolane hydrochloride (Traut's reagent), albumin from bovine serum (BSA), and Tf were purchased from Sigma-Aldrich (St. Louis, MO, USA). Methoxy-PEG5000-b-PCL15000 was purchased from Daigang Biomaterial Co., Ltd. (Jinan, Shandong, China). NH2-PEG5800-b-PCL19000 was purchased from Polymer Source Inc. (Montreal, Quebec, Canada). Other chemicals and reagents used in the study were of analytical grade.
All cell lines were preserved in our lab. BALB/c nude mice (male, 4 to 6 weeks) were purchased from Hunan Slake Jingda Experimental Animal Co. Ltd., Changsha, China (Animal Qualification Certification No. 43004700000595). All animal studies were approved by the Animal Experimentation Ethics Committee of College of Life Science and Technology, Huazhong University of Science and Technology and carried out in compliance with the guidelines approved by the Science and Technology Department of Hubei Province.
All cell lines were preserved in our lab. BALB/c nude mice (male, 4 to 6 weeks) were purchased from Hunan Slake Jingda Experimental Animal Co. Ltd., Changsha, China (Animal Qualification Certification No. 43004700000595). All animal studies were approved by the Animal Experimentation Ethics Committee of College of Life Science and Technology, Huazhong University of Science and Technology and carried out in compliance with the guidelines approved by the Science and Technology Department of Hubei Province.
9
Isolation and Characterization of Human Satellite Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human satellite cells were isolated from muscle biopsies as reported by Cardani et al.43 Myoblasts were grown in HAM’s F10 medium (Sigma-Aldrich) supplemented with 15% FBS (Euroclone), 0.5 mg/mL albumin from bovine serum (BSA, Sigma-Aldrich), 0.5 mg/mL fetuin (Sigma-Aldrich), 0.39 µg/mL dexamethasone, 10 ng/mL epidermal growth factor, 0.05 mg/mL insulin, 3 mg/mL glucose, 100 U/mL penicillin and 100 µg/mL streptomycin (proliferative medium). For this study, cells from DM and control patients were plated at a density of 85,000 cells per 60 mm dishes or 50,000 cells per 35 mm dishes. Myogenic purity was evaluated by immunofluorescence using desmin as marker (see below). The percentage of desmin positive myoblasts was calculated as the number of positive cells vs the total number of cells observed. All starting cell populations used in this study had a myogenic purity higher than 80%. Moreover, the myogenic purity was monitored every 5 passages (7 days per passage for a total of 35 days) until senescence was reached. At least 100 cells were counted in at least 10 different optical fields randomly chosen.
10
Brucella melitensis Rev1 Vaccine Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Brucella melitensis Rev1 vaccine strain (Biovar 1) was obtained from Dollvet (Veterinary Vaccine, Medicine, Biological Article Generation Industry and Trade Inc., Sanliurfa/Turkey). Escherichia coli One Shot® Mach1T1R™ served for cloning and plasmid amplification. E. coli BL21 (DE3) served as the expression host. Bacterial strains were stored at –80°C until the time of use. Champion™ pET SUMO Protein Expression System was purchased from Invitrogen, USA. GenElute™ Plasmid Miniprep Kit, Lysozyme from chicken egg white, dNTP Mix, Taq DNA Polymerase, ampicillin, kanamycin sulphate, and albumin from bovine serum (BSA) were purchased from Sigma-Aldrich, USA. Antigen of RBPT was obtained from Pendik Veterinary Control and Research Institute. Ni-NTA colon was purchased from Thermo (USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!