The adhesion to the intestinal epithelium of the eight strains from Andean food origin, together with the two probiotics strains used as reference, was tested on the cell line HT29 (ECACC 91072201, European Collection of Cell Cultures, UK), according to procedures previously described [14] (link). Briefly, bacterial cultures (10 mL) were washed twice with PBS and resuspended in 1 mL (at 10 8 CFU/mL) of the complete (without antibiotics) McCoy's medium (Sigma) specific for this cell line. Then, 13 ± 1-day-old HT29 cell monolayers, seed on 12-well plates, were gently washed with Dulbecco PBS (Sigma) before adding the bacterial suspensions. Monolayers, with bacteria-HT29 cell ratio around 10:1, were incubated for 1 h at 37 °C/5% CO 2 . Supernatants were removed and gently washed twice with Dulbecco PBS before adding 0.5 mL of 0.25% EDTA-trypsin (Sigma). Plates were incubated under same conditions for 10-15 min, to favour the release of the cells from the monolayer, and then 0.5 mL of complete-McCoy's medium was added. Bacterial counts of the initial cells added as well as those adhered to HT29 cells were enumerated by plating in agar-MRS. The percentage of adhesion was calculated as the CFU adhered bacteria divided by the CFU of added bacteria. This procedure was performed with triplicated cultures of each lactobacilli strain.
Dulbecco pbs
Manufactured by Merck Group
Sourced in France, United States, Germany
Dulbecco's Phosphate Buffered Saline (Dulbecco PBS) is a sterile, isotonic buffer solution commonly used in various laboratory applications. It is a balanced salt solution that maintains the pH and osmolality of cell culture media, enabling the preservation of cellular integrity and function. Dulbecco PBS is a versatile reagent widely used in cell biology, immunology, and other life science research fields.
Automatically generated - may contain errors
Lab products found in correlation
Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions)
Mccoy s medium, by Merck Group (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Superdex 75, by GE Healthcare (1 mentions)
Reduced β nicotinamide adenine dinucleotide phosphate nadph, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Methyl β cyclodextrin mβcd, by Merck Group (1 mentions)
Ni sepharose, by GE Healthcare (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Jsm 840 sem, by JEOL (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Perm wash buffer, by BD (1 mentions)
Imipramine, by Merck Group (1 mentions)
Coomassie brilliant blue g 250, by Serva Electrophoresis (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Carboxy h2dcfda, by Thermo Fisher Scientific (1 mentions)
Glutaraldehyde, by Ted Pella (1 mentions)
Mci buffer l 8800 ph 1 4, by Fujifilm (1 mentions)
Trypsin, by Promega (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by MP Biomedicals (1 mentions)
7 protocols using dulbecco pbs
1
Adhesion of Andean Lactobacilli to Intestinal Cells
2
Equine Cytochrome c Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Equine heart cytochrome c (cyt c), arachidonic acid (cis-AA), phenylmethanesulfonyl fluoride (PMSF), isopropylthiogalatoside (IPTG), cholesterol, Dulbecco PBS and methyl-β-cyclodextrin (MβCD) were from Sigma (Saint-Quentin Fallavier, France). Reduced β-nicotinamide adenine dinucleotide phosphate (NADPH) was from Acros. Ni-sepharose, superdex 75 and Ficoll-Paque Plus was from GE Healthcare, France.
3
Scanning Electron Microscopy of Biofilms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formed biofilms of MSSA and MRSA were fixed in 2.5 % (vol/vol) glutaraldehyde (Sigma, USA) in Dulbecco PBS (PH 7.2) for 1.5 h, rinsed with PBS, and then dehydrated through an ethanol series. Samples were dried and gold–palladium coated. SEM examinations were made on a JSM-840 SEM (JEOL Ltd., Tokyo, Japan) [27 (link)]. SEM was used in the current study for visualizing the formed biofilms and most importantly for assessing visually the type of the formed biofilms whether polysaccharide- or protein- based biofilm. The IPA-based biofilm appears usually with rough crystallized surface while FnBPA-based biofilm appears as smooth non-crystallized surface.
4
Apoptosis Induction Assay for PBMC and Melanoma
5
Murine Bronchoalveolar Lavage and Lung Tissue Collection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In some studies, following pulmonary function assessment, anesthetized mice, were lavaged through an 18-g tracheal cannula with 1 mL of room temperature Dulbecco PBS (Sigma-Aldrich). Cells were manually counted immediately in white blood cell stain (0.2 mg/mL of crystal violet in 2% acetic acid) by using a hemocytometer. The bronchoalveolar lavage fluid (BALF) was centrifuged at 400 g for 10 minutes at room temperature, and cell-free supernatants were frozen at –80°C until analysis of total protein content by bicinchoninic acid assay (Pierce Biotechnology). Cell pellets were resuspended in saline and mounted on slides by cytospin (100,000 cells per slide) for hematoxylin and eosin staining and differential analysis. Following collection of the BALF, the lungs were dissected, ground to a fine powder using a liquid nitrogen–chilled mortar and pestle, and stored at –80°C until analysis.
6
Murine Bronchoalveolar Lavage Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In some studies, following pulmonary function assessment, anesthetized mice, were lavaged through an 18-g tracheal cannula with 1 mL of room temperature Dulbecco PBS (Sigma-Aldrich). Cells were manually counted immediately in white blood cell stain (0.2 mg/mL of crystal violet in 2% acetic acid) by using a hemocytometer. The bronchoalveolar lavage fluid (BALF) was centrifuged at 400 g for 10 minutes at room temperature, and cell-free supernatants were frozen at −80°C until analysis of total protein content by bicinchoninic acid assay (Pierce Biotechnology). Cell pellets were resuspended in saline and mounted on slides by cytospin (100,000 cells per slide) for hematoxylin and eosin staining and differential analysis. Following collection of the BALF, the lungs were dissected, ground to a fine powder using a liquid nitrogen–chilled mortar and pestle, and stored at −80°C until analysis.
7
Bicarbonate-free Hank's solution analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bicarbonate-free Hank’s solution, Ca2+-free Dulbecco PBS, imipramine, LPS (lipopolysaccharide from Salmonella enterica serovar Typhimurium) and E64 were obtained from Sigma (Steinheim, Germany). Ficoll-Paque was obtained from Pharmacia (Uppsala, Sweden). Fibronectin was from Calbiochem (La Jolla, United States). Coomassie Brilliant Blue G-250 was obtained from Serva, PMSF from MP Biomedical, trypan blue from Fluka AG, glutaraldehyde from Ted Pella. Trypsin was from Promega, carboxy-H2DCF-DA from Molecular probe, United States. Analytical chromatography conditions: eluent MCI Buffer L-8800-PH-1–4 and ninhydrin coloring solution kit for Hitachi 29970501 (Wako Chemicals GmbH, United States).
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!