Cells were incubated for 24 h in complete media supplemented with 1 µg/ml doxycycline and 50 µM biotin. After three PBS washes, cells (for small-scale analysis, <107; for large scale analysis, 4 × 107) were lysed at 25°C in 1 ml lysis buffer (50 mM Tris, pH 7.4, 500 mM NaCl, 0.4% SDS, 5 mM EDTA, 1 mM DTT, and 1x Complete protease inhibitor [Roche]) and sonicated. Triton X-100 was added to 2% final concentration. After further sonication, an equal volume of 4°C 50 mM Tris (pH 7.4) was added before additional sonication (subsequent steps at 4°C) and centrifugation at 16,000 relative centrifugal force. Supernatants were incubated with 600 µl Dynabeads (MyOne Steptavadin C1; Invitrogen) overnight. Beads were collected and washed twice for 8 min at 25°C (all subsequent steps at 25°C) in 1 ml wash buffer 1 (2% SDS in dH2O). This was repeated once with wash buffer 2 (0.1% deoxycholate, 1% Triton X-100, 500 mM NaCl, 1 mM EDTA, and 50 mM Hepes, pH 7.5), once with wash buffer 3 (250 mM LiCl, 0.5% NP-40, 0.5% deoxycholate, 1 mM EDTA, and 10 mM Tris, pH 8.1) and twice with wash buffer 4 (50 mM Tris, pH 7.4, and 50 mM NaCl). 10% of the sample was reserved for Western blot analysis. Bound proteins were removed from the magnetic beads with 50 µl of Laemmli SDS-sample buffer saturated with biotin at 98°C. For the larger scale preparation, 90% of the sample to be analyzed by mass spectrometry was washed twice in 50 mM NH4HCO3.
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Deoxycholate
Deoxycholate
Deoxycholic acid, also known as deoxycholate, is a secondary bile acid produced in the liver as a product of cholesterol metabolism.
It serves as an emulsifier, aiding in the digestion and absorption of fats.
Deoxycholate has a wide range of biological activities, including roles in lipid metabolism, cell signaling, and inflammation.
It is commonly used in biochemical and cell biology applications, such as cell lysis and protein extraction.
Reserchers can use PubCompare.ai's AI-powered optimizatioon to locate the best protocols and products for working with deoxychloate and other biomolecules, supporting more reproducible and efficient research.
It serves as an emulsifier, aiding in the digestion and absorption of fats.
Deoxycholate has a wide range of biological activities, including roles in lipid metabolism, cell signaling, and inflammation.
It is commonly used in biochemical and cell biology applications, such as cell lysis and protein extraction.
Reserchers can use PubCompare.ai's AI-powered optimizatioon to locate the best protocols and products for working with deoxychloate and other biomolecules, supporting more reproducible and efficient research.
Most cited protocols related to «Deoxycholate»
Biotin
Buffers
Cells
Centrifugation
Deoxycholate
Doxycycline
Edetic Acid
HEPES
Laemmli buffer
Mass Spectrometry
Nonidet P-40
Protease Inhibitors
Proteins
Sodium Chloride
Triton X-100
Tromethamine
Western Blot
Acetone
Acid Hybridizations, Nucleic
Biotin
Bistris
Buffers
Cells
Cold Temperature
Deoxycholate
Edetic Acid
Formaldehyde
Gels
Glycine
HEPES
Hyperostosis, Diffuse Idiopathic Skeletal
Mass Spectrometry
Pellets, Drug
Proteins
Ribonucleases
SDS-PAGE
Sodium Chloride
sodium lauroyl sarcosinate
Vertebral Column
Western Blot
Bath
BP 400
Buffers
Cell Extracts
Cell Lines
Cell Nucleus
Cells
Chromatin
Cytosol
Deoxycholate
Dietary Fiber
DNA Chips
Edetic Acid
Egtazic Acid
Formaldehyde
Glycerin
HEPES
Ice
Igepal CA-630
isolation
N-lauroylsarcosine
Nonidet P-40
Polypropylenes
Protease Inhibitors
Proteins
Sepharose
Sodium Chloride
Tissues
Triton X-100
Tromethamine
Trypan Blue
Anabolism
beta-Actin
CD4 Positive T Lymphocytes
CD8-Positive T-Lymphocytes
CDK9 protein, human
Cells
Cyclin T1
Deoxycholate
DNA, Complementary
Nitrocellulose
Oligonucleotide Primers
Perforin
PRDM1 protein, human
Protease Inhibitors
Proteins
SDS-PAGE
Sodium Chloride
T-Lymphocyte
TBX21 protein, human
Tissue, Membrane
Triton X-100
Tromethamine
Western Blot
ChIP experiments were performed as previously described previously12 (link), with some changes. S2 Drosophila cultured cells were fixed in formaldehyde (Sigma) at a final concentration of 1.8% for 10 min. After several washes, the cells were homogenized using a dounce homogenizer, pelleted, resuspended in cold buffer, and SDS added to a final concentration of 1%. Cells were again pelleted, washed and finally resuspended at a final concentration of 1×108 nuclei/ml, with 0.1% SDS. Cells were sonicated using a Bioruptor sonicator. All lysates were combined, after which Triton-X 100 and Deoxycholate were added. After centrifugation, the final supernatant contained soluble chromatin. Input chromatin was treated with RNase, followed by Proteinase K, and crosslinking was reversed. The average size of the DNA fragments was 400–1000 bp. For ChIP, chromatin was pre-cleared by incubating with Protein A Sepharose beads. After the beads were removed, chromatin was incubated with the antibody for immunoprecipitation, followed by addition of Protein A Sepharose beads. After washing, sample attached to beads was treated with RNase A, followed by Proteinase K, and crosslinking was reversed. Half of each ChIP sample and 50 ng of Input DNA were amplified using a WGA Kit (Sigma # WGA2). Samples were purified using a QIAquick PCR purification column (QIAGEN). The amplified DNA was fragmented using RNAse-free DNaseI, after which the peak of bulk DNA was at 50–100 bp. The fragmented DNA library was labeled with biotin by a terminal deoxynucleotidyl transferase reaction, and hybridization cocktail was added. Genomic DNA Tiling Arrays v2.0 (Affymetrix) were pre-hybridized and then hybridized to ChIP sample or Input DNA for 18 hours, followed by washing and staining in a fluidics station EukGE-WS2v4 (Affymetrix). Enrichment P-values are calculated using a sliding window (default size 1 kb) moved in steps across the genome (default step size 30 bp). A P-value enrichment score is calculated at each step using a one-sided T-test on the M-values of probes that fall within the window. To capture both significant enrichment and significant depletion, P-values for enrichment test (ePv) and depletion test (dPv) are calculated, and the score is given as −log10(min(ePv, dPv)). The score is multiplied by −1 if dPv was smaller than ePv.
Acid Hybridizations, Nucleic
Biotin
BP 100
Buffers
Cell Nucleus
Cells
Centrifugation
Chromatin
Cold Temperature
Cultured Cells
Deoxycholate
Dietary Fiber
DNA Chips
DNA Library
DNA Nucleotidylexotransferase
Drosophila
Endopeptidase K
Endoribonucleases
Formaldehyde
Genome
Immunoglobulins
Immunoprecipitation
Staphylococcal protein A-sepharose
Triton X-100
Most recents protocols related to «Deoxycholate»
This study was conducted in Damietta Governorate on the Egyptian Mediterranean coast (northern east Nile Delta), Egypt through the period from October 2021 to March 2022. A total of 200 cloacal swabs were collected from migratory and broiler chicken birds. Broiler chickens were selected from poultry farms and live bird markets near which the migratory birds were hunted at the similar time points. One hundred samples were obtained from migratory birds and 100 from broiler chickens; 50 from 5 poultry farms (10 for each farm) with deep litter system and 50 from 3 live bird markets located in different regions inside Damietta Governorate. Five broiler poultry farms were chosen on the basis of their owners’ willingness to permit the samples collection. Broiler chicken birds from the farms and live bird markets were selected randomly. The map of Damietta Governorate was constructed to highlight the location of the selected broiler chicken farms and live bird markets in relation to the rest of Damietta (Supplementary Fig. 9 ). The migratory birds that were found near to the examined farms and live bird markets were trapped by net traps, sampled, marked (to ensure that each bird was only sampled once) and photographed to detect its species. The cotton swabs were aseptically collected on 2 ml of Bolton broth (Oxoid, UK) then labeled and transported within 1 h in an ice box at 4 °C to the Reference Laboratory for Veterinary Quality control on Poultry production to perform further examinations. All samples were incubated at 42 °C for 48 h under microaerophilic conditions. Isolation and identification of Campylobacter spp.
Each enriched sample was streaked onto modified charcoal cefoperazone deoxycholate agar (Oxoid, UK) with antibiotic solution (cefoperazone sodium salt; 0.032 g, amphotericin B; 0.01 g and water; 5 ml) and incubated at 42 °C for 48 h. The suspected colonies were identified by morphological characteristics and Gram staining [45 ]. The suspected isolates were subjected to standard biochemical procedures, including tests for hippurate, acetate hydrolysis and catalase [46 ].
Each enriched sample was streaked onto modified charcoal cefoperazone deoxycholate agar (Oxoid, UK) with antibiotic solution (cefoperazone sodium salt; 0.032 g, amphotericin B; 0.01 g and water; 5 ml) and incubated at 42 °C for 48 h. The suspected colonies were identified by morphological characteristics and Gram staining [45 ]. The suspected isolates were subjected to standard biochemical procedures, including tests for hippurate, acetate hydrolysis and catalase [46 ].
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Acetate
Agar
Amphotericin B
Antibiotics
Aves
Campylobacter
Catalase
Cefoperazone
Charcoal
Chickens
Deoxycholate
Enterobacter
Fowls, Domestic
Gossypium
Hartnup Disease
hippurate
Hydrolysis
isolation
Physical Examination
Sodium, Cefoperazone
Specimen Collection
The transfected control Caco2 cells were lysed in RIPA buffer (150 mM NaCl; 50 mM Tris-Cl, pH 8; 1% NP-40; 0.5% deoxycholate; 0.1% SDS) (Beyotime, Shanghai, China) with Protease Inhibitor Cocktail (100×) (Thermo Scientific, MA, USA). The resulting samples were separated using 10% SDS-PAGE (Invitrogen, Carlsbad, CA, USA) and transferred to polyvinylidene fluoride (PVDF) membranes. After blocking for 1 h in 5% non-fat milk in TBST at 25°C, the membranes were incubated overnight at 4 °C with specific primary antibodies, including SMARCE1 (Abcam, Cambridge, UK, 1:1000), tfec (Santa Cruz, USA,1:500), and β-tubulin (Sigma, USA, 1:10,000). Primary antibody binding was visualized using horseradish peroxidase-conjugated goat anti-rabbit or anti-mouse IgG (Sigma, USA, 1:10,000) for 1 h at 25°C. Signal intensities were measured using a Chemiluminescent HRP substrate (Yamei, Shanghai, China) and imaged using ImageJ v1.8.0 (NIH, Bethesda, MD, USA).
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anti-IgG
Antibodies
Buffers
Caco-2 Cells
Deoxycholate
Goat
Immunoglobulins
Milk, Cow's
Mus
Nonidet P-40
polyvinylidene fluoride
Protease Inhibitors
Rabbits
Radioimmunoprecipitation Assay
SDS-PAGE
SMARCE1 protein, human
Sodium Chloride
Tissue, Membrane
Tromethamine
Tubulin
Cells were lysed with ice cold lysis buffer buffer (25 mM HEPES, pH 7.4, 150 mM NaCl, 1% NP-40, 0.25% Na Deoxycholate, 10% glycerol) supplemented with protease and phosphatase inhibitor cocktails (539134; MilliporeSigma; BP-479; Boston BioProducts), and incubated on ice for 5 min. Cellular debris was pelleted out by centrifugation at 21,000 g for 15 min at 4°C. Protein concentration was determined using a Bradford assay compared to BSA protein standards. Supernatant was mixed with Laemmli buffer and boiled at 95°C for 5 min. To determine protein expression, 20–30 µg of protein lysate was loaded per well. For immunoprecipitation blots, 5% of input lysate and entire IP eluate was loaded. Samples were resolved using SDS-PAGE and 10% acrylamide gels. Resolved samples were transferred to nitrocellulose membranes (926-31090; LI-COR), and membranes were blocked in Intercept Blocking Buffer (927-70001; LI-COR) for 1 h rocking at room temperature. Membranes were probed overnight at 4°C with primary antibodies diluted in blocking buffer and 0.2% Tween-20. After washing in TBS with 0.1% Tween-20, membranes were probed with near-infrared fluorescent secondary antibodies diluted in blocking buffer and 0.2% Tween-20 for 1 h at room temperature. Membranes were scanned using an Odyssey CLX imager (LI-COR) and quantified in Image Studio Lite (LI-COR). Antibodies were as follows: rabbit polyclonal anti-EVL (a gift from Frank Gertler, 1:1,000), rabbit polyclonal anti-ENAH (HPA028696; Sigma Prestige, RRID:AB_10611249; 1:250), rabbit monoclonal anti-VASP (#3132; Cell Signaling, RRID:AB_2213393; 1:500), rabbit polyclonal anti-MTSS1 (PA5-23200; Thermo Fisher Scientific, RRID:AB_2540726; 1:500) rabbit polyclonal anti-FLAG (20543-1; ProteinTech Group, RRID:AB_11232216; 1:1,000), rabbit polyclonal anti-GFP (66002-1; ProteinTech Group, RRID:AB_11182611; 1:1,000), mouse monoclonal anti-actin (66009-1; ProteinTech Group, RRID:AB_2687938; 1:5,000), goat anti-mouse Alexa Fluor 680 (#A-21057; Thermo Fisher Scientific, RRID:AB_2535723; 1:20,000) and goat anti-rabbit Alexa Fluor 790 secondary antibodies (#A-11367; Thermo Fisher Scientific, RRID:AB_2534141; 1:20,000).
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Acrylamide
Actins
Anti-Antibodies
Antibodies
Biological Assay
Buffers
Cells
Centrifugation
Cold Temperature
Deoxycholate
Fluorescent Antibody Technique
Gels
Glycerin
Goat
HEPES
Immunoprecipitation
Laemmli buffer
Mus
Nitrocellulose
Nonidet P-40
Peptide Hydrolases
Phosphoric Monoester Hydrolases
Prestige dental material
Proteins
Rabbits
SDS-PAGE
Sodium Chloride
Tissue, Membrane
Tween 20
VASP protein, human
Chicken skins were randomly collected from Koch Food Company (USA) and sliced into 10 cm × 10 cm samples. To minimize contamination, chicken skin was washed with 200 ppm chlorine solution (Sigma-Aldrich Co.) and sterilized DW. Then, 200 ml of the Salmonella cocktail was inoculated onto the chicken at concentrations ranging from 101 to 103 CFU/100 cm2. An equal amount of PBS was added onto other chicken skins as negative controls. The inoculated chicken skins were dried under a biosafety cabinet for bacterial attachment and placed in an Erlenmeyer flask prior to further incubation in a refrigerator (4°C) for 48 h. Next, 100 ml of brain heart infusion (BHI, EMD Science, Germany) or brilliant green (BG, Difco Laboratories Inc.) broth was added to each flask and incubated at 37°C in an orbital shaker at 250 rpm. Then, 100 μl of sample was collected from BHI and BG broths at 0, 2, 4, and 6 h, and the resuscitated bacterial population was measured using xylose lysine deoxycholate agar (Difco Laboratories Inc.) and recorded as log CFU/chicken for comparison. Subsequently, 20 ml of samples were obtained from both broths and washed 3 times by centrifugation at 4,000 ×g for 20 min. After resuspending with 1 ml of PBS, 100 μl of Salmonella suspension was used for ELISA and GB-LMIS, as described in the previous section. The results are expressed as log CFU/chicken for the comparison.
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Agar
Bacteria
Brain
brilliant green
Centrifugation
Chickens
Chlorine
Deoxycholate
Enzyme-Linked Immunosorbent Assay
Food
Heart
Keratosis pilaris
Lysine
Salmonella
Xylose
Method described previously (Nacke et al., 2021 (link)); briefly, PC3 cells were incubated on 120-mm plates for 48 h at 37°C, 5% CO2. Cells were then lysed on ice in pulldown-lysis buffer (50 mM Tris, 100 mM NaCl, 2 mM MgCl2, 0.1% SDS, 0.5% Na-deoxycholate, 1% Triton X-100, 10% glycerol). Lysates were syringed 5× using a 25-27G needle and centrifuged at 4°C 14,000 g for 1 min. Spin columns were equilibrated with 50 μl of Glutathione Agarose resin and washed with pulldown-column wash buffer (1:1 pulldown-lysis buffer and 1×TBS). 80 μg of GST-GGA3-GAT recombinant fusion protein was immobilized on the agarose resin by incubation at 4°C with gentle rocking. After 1 h, 1 mg of each lysate was added onto spin columns and incubated again at 4°C for 2 h with rocking. Pulldown wash buffer (50 mM Tris, 100 mM NaCl, 2 mM MgCl2, 1% NP-40, 10% glycerol) was used to wash unbound proteins off the column. 60 μl of pulldown-elution buffer (10 mM Glutathione in 1×TBS) was added to each spin column and incubated for 5 min at room temperature. Eluted protein was collected at 1,250 g for 1 min and samples prepared for SDS-PAGE and immunoblotting as described above. n = 3 experimental replicates for ARF3 Chimeras and n = 5 for PSD KD. Data is presented as GGA3 binding normalized to ARF3 or ARF6 levels for each experiment. Values are mean ± SEM.
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ARF6 protein, human
Buffers
Cells
Chimera
Deoxycholate
Glutathione
Glycerin
Magnesium Chloride
Needles
Nonidet P-40
PC 3 Cell Line
Proteins
Recombinant Fusion Proteins
Resins, Plant
SDS-PAGE
Sepharose
Sodium Chloride
Triton X-100
Tromethamine
Top products related to «Deoxycholate»
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Protease inhibitor cocktail is a laboratory reagent used to inhibit the activity of proteases, which are enzymes that break down proteins. It is commonly used in protein extraction and purification procedures to prevent protein degradation.
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The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
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The Complete Protease Inhibitor Cocktail is a laboratory product designed to inhibit a broad spectrum of proteases. It is a concentrated solution containing a mixture of protease inhibitors effective against a variety of protease classes. This product is intended to be used in research applications to preserve the integrity of target proteins by preventing their degradation by proteolytic enzymes.
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The Pierce BCA Protein Assay Kit is a colorimetric-based method for the quantification of total protein in a sample. It utilizes the bicinchoninic acid (BCA) reaction, where proteins reduce Cu2+ to Cu+ in an alkaline environment, and the resulting purple-colored reaction is measured spectrophotometrically.
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β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.
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Nitrocellulose membranes are a type of laboratory equipment designed for use in protein detection and analysis techniques. These membranes serve as a support matrix for the immobilization of proteins, enabling various downstream applications such as Western blotting, dot blotting, and immunodetection.
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The BCA Protein Assay Kit is a colorimetric detection and quantification method for total protein concentration. It utilizes bicinchoninic acid (BCA) for the colorimetric detection and quantification of total protein. The assay is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium, with the chelation of BCA with the Cu1+ ion resulting in a purple-colored reaction product that exhibits a strong absorbance at 562 nm, which is proportional to the amount of protein present in the sample.
Sourced in United States, Italy
Deoxycholate is a laboratory reagent commonly used in biochemical and molecular biology applications. It is a bile salt derived from cholic acid and functions as a detergent, facilitating the solubilization and extraction of proteins, nucleic acids, and other biomolecules from cellular and subcellular samples. The core function of deoxycholate is to disrupt cell membranes and solubilize hydrophobic compounds, enabling their isolation and analysis.
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Protease inhibitors are a class of laboratory equipment used in the field of biochemistry and molecular biology. These inhibitors are designed to specifically target and inactivate proteases, which are enzymes that break down proteins. Protease inhibitors play a crucial role in various experimental and analytical procedures, such as protein extraction, purification, and stabilization.
More about "Deoxycholate"
Deoxycholic acid, also known as deoxycholate, is a secondary bile acid produced in the liver as a byproduct of cholesterol metabolism.
It plays a crucial role as an emulsifier, aiding in the digestion and absorption of fats.
Deoxycholate has a wide range of biological activities, including involvement in lipid metabolism, cell signaling, and inflammation.
It is commonly utilized in biochemical and cell biology applications, such as cell lysis and protein extraction.
Researchers can leverage PubCompare.ai's AI-powered optimization to locate the most effective protocols and products for working with deoxycholate and other biomolecules, supporting more reproducible and efficient research.
This includes finding the best solutions for protease inhibitor cocktails, PVDF and nitrocellulose membranes, BCA protein assays, and β-actin detection.
PubCompare.ai's platform allows you to easily identify the most relevant protocols from literature, preprints, and patents, while its AI-driven comparisons help you pinpoint the most effective products and methods.
This can lead to smarter, more efficeint research, ultimately advancing your studies on deoxycholate and other key biomolecules.
It plays a crucial role as an emulsifier, aiding in the digestion and absorption of fats.
Deoxycholate has a wide range of biological activities, including involvement in lipid metabolism, cell signaling, and inflammation.
It is commonly utilized in biochemical and cell biology applications, such as cell lysis and protein extraction.
Researchers can leverage PubCompare.ai's AI-powered optimization to locate the most effective protocols and products for working with deoxycholate and other biomolecules, supporting more reproducible and efficient research.
This includes finding the best solutions for protease inhibitor cocktails, PVDF and nitrocellulose membranes, BCA protein assays, and β-actin detection.
PubCompare.ai's platform allows you to easily identify the most relevant protocols from literature, preprints, and patents, while its AI-driven comparisons help you pinpoint the most effective products and methods.
This can lead to smarter, more efficeint research, ultimately advancing your studies on deoxycholate and other key biomolecules.