Human recombinant IL-1α and human recombinant IGF-1 were purchased from PeproTech (Rocky Hill, NJ). Dulbecco’s modification of Eagle’s medium (DMEM) was purchased from Corning (Corning, NY). Trypsin-EDTA phenol red, HEPES buffer, non-essential amino acids (NEAA), and penicillin streptomycin antibiotic-antimycotic (PSA) were purchased from Gibco (Carlsbad, CA). Ascorbic acid (AA) and L-proline were from Fisher Bioreagents (Pittsburgh, PA). Proteinase-K was purchased from Roche Diagnostics (Risch-Rotkreuz, Switzerland). NHS-PEG2-Maleimide and Protease Inhibitor Mini Tablets were from Thermo Scientific Pierce (Rockford, IL). Propidium iodide (PI) was obtained from Thermofisher Acros Organics (Geel, Belgium). Tris-HCl buffer was purchased from Invitrogen (Grand Island, NY). Chondroitinase ABC, Resazurin sodium salt, Griess reagent, fluorescein diacetate (FDA), and other salts and reagents were purchased from Sigma (St. Louis, MO).
Tris hcl buffer
Manufactured by Thermo Fisher Scientific
Sourced in United States
Tris-HCl buffer is a common buffer solution used in various laboratory applications. It is composed of tris(hydroxymethyl)aminomethane (Tris) and hydrochloric acid (HCl), which together provide a buffering system to maintain a specific pH range. The primary function of Tris-HCl buffer is to maintain a stable pH environment for experiments and reactions.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (9 mentions)
Acetonitrile, by Thermo Fisher Scientific (6 mentions)
Methanol, by Thermo Fisher Scientific (4 mentions)
Deep well 1 ml 96 well plate protein lobind, by Eppendorf (2 mentions)
Recombinant mouse hexb protein his tag, by MyBioSource (2 mentions)
Enzyme activity assay kit for β n acetyl glucosaminidase, by Merck Group (2 mentions)
Recombinant nag enzyme, by New England Biolabs (2 mentions)
Instant pc kit, by Agilent Technologies (2 mentions)
Dithiothreitol (dtt), by Thermo Fisher Scientific (2 mentions)
Iodoacetamide iam, by Thermo Fisher Scientific (2 mentions)
Sequencing grade modified trypsin, by Promega (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
T4 dna ligase, by Thermo Fisher Scientific (2 mentions)
Fluorescein diacetate, by Merck Group (1 mentions)
Chondroitinase abc, by Merck Group (1 mentions)
Ascorbic acid, by Thermo Fisher Scientific (1 mentions)
Trypsin edta phenol red, by Thermo Fisher Scientific (1 mentions)
L proline, by Thermo Fisher Scientific (1 mentions)
Recombinant human il 1α, by Thermo Fisher Scientific (1 mentions)
Recombinant human igf 1, by Thermo Fisher Scientific (1 mentions)
15 protocols using tris hcl buffer
1
Chondrocyte Differentiation Induction Protocol
2
RNA Transfection and Cytotoxicity Assay
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All oligonucleotides were purchased from Integrated DNA Technologies (IDT, Skokie, IL, USA). T4 ligase was obtained from Promega (Madison, WI, USA). T7 RNA polymerase and ribonucleotide triphosphates (rNTPs) were purchased from New England BioLabs (NEB, Ipswich, MA, USA). ZebaTM spin desalting columns were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Cyanine 5-UTP was purchased from Enzo Life Sciences (Farmingdale, NY, USA). Mica and Lacey Formvar/carbon-coated copper grids (01883-F) were obtained from Ted Pella (Redding, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM), penicillin/streptomycin (P/S), fetal bovine serum (FBS), and Dulbecco’s phosphate buffered saline (DPBS) were purchased from Gibco (Waltham, MA, USA). The StemfectTM RNA Transfection Kit was purchased from Stemgent (Houston, TX, USA). CelLytic M and MgCl2 solution were purchased from Sigma-Aldrich (Saint Louis, MI, USA). A Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Laboratories (Kumamoto, Japan). Tris-HCl buffer was purchased from Invitrogen (Waltham, MA, USA).
3
Proteinase Extraction and Purification
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Extraction of proteinase was performed as previously described by Villegas et al. (2015) (link) with slight modifications. Lactococcus and Lactobacillus cells from the exponential growth phase were harvested by centrifugation at 8000×g for 10 min at 4 °C, washed twice with 0.85 % (m/V) saline supplemented with 10 mM CaCl 2 (Gram-Mol, Zagreb, Croatia), and resuspended to in 20 mM Tris-HCl buffer of pH 7.5 (Invitrogen, Carlsbad, USA) containing 5 mM CaCl 2 . Samples were incubated for 30 min at room temperature and then centrifuged at 8000 × g for 15 min. The supernatants of protein extracts were stored at 4 °C, and the precipitated protein pellets were washed, resuspended in 20 mM Tris-HCl buffer and stored at 4 °C. The protein concentration was measured by a BioSpec-nanospectrophotometer (Shimadzu, Japan). Supernatants of protein extracts and protein pellet fractions were subjected to proteolytic activity and SDS-PAGE analyses.
In order to further concentrate and purify proteinase, ultrafiltration of supernatants of 7 selected strains, obtained after proteinase extractions, was performed by filtration through a 100-kDa membrane (100,000 MWCO, Amicon, Millipore) at 4000 x g during 10 min, according to the manufacturer's instruction, yielding a partially purified proteinase sample.
In order to further concentrate and purify proteinase, ultrafiltration of supernatants of 7 selected strains, obtained after proteinase extractions, was performed by filtration through a 100-kDa membrane (100,000 MWCO, Amicon, Millipore) at 4000 x g during 10 min, according to the manufacturer's instruction, yielding a partially purified proteinase sample.
4
Screening Amoxicillin-Resistant Isolates for Carbapenemase Production
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Up to 100 randomly selected amoxicillin-resistant colonies per sample were screened for carbapenemase production by CarbaNP test (Dortet et al., 2014 (link)). Briefly, cells were lysed in 100 μl of Tris-HCl buffer (Thermo Scientific, Rockford, Il, USA) and the lysate was mixed with 100 μl of phenol red solution containing 6 mg/ml imipenem/cilastatin (Fresenius Kabi, Bad Homburg, Germany). Phenol red solution without imipenem was included in the test as a negative control. After incubating at 37°C for a maximum of 2 h, red to orange/yellow color shift in the test vial and no color change in the negative control were interpreted as imipenem hydrolysis. Plasmid inserts of the carbapenemase-producing clones were sequenced using the primers described in Table 2 . Sequences displaying less than 70% amino acid sequence identity to known MBLs were defined as new MBLs, as suggested by Cornaglia et al. (2007 (link)).
5
Evaluation of Bioactive Compounds from Korean Persimmon
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D. morbifera was grown and harvested in Wando, Korea. Methanol and acetonitrile were purchased from Honeywell Burdic & Jackson (Morristown, NJ, USA). Formic acid, sodium carbonate (Na2CO3), and aluminum chloride hexahydrate (AlCl3·6H2O) were purchased from Daejung Chemicals & Metals Co., Ltd. (Siheung-si, Korea). EZ-cytox solution was purchased from DoGenBio (Guro-gu, Korea). Tris-HCl buffer, PBS, and Tris solution buffer were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Folin–Denis reagent, kojic acid, elastase, mushroom tyrosinase, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), Cell Proliferation Kit I (MTT), DMSO, L-ascorbic acid, gallic acid, and quercetin were purchased from Sigma Aldrich (St. Louis, MO, USA)
6
Therapeutic mAb Immunoglobulin (IgG) Characterization
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We purchased the therapeutic mAb immunoglobulin (IgG) gamma 1 drugs adalimumab and rituximab from Eisai Co., Ltd. (Tokyo, Japan) and Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan), respectively. In addition, we purchased Tris-HCl buffer (pH 8.0), 10% trifluoroacetic acid, 0.1% formic acid, acetonitrile 0.1% formic acid and 7 K Dialysis Casettes from Thermo Fisher Scientific (San Jose, CA, USA). We obtained 8 M guanidine hydrochloride from Sigma-Aldrich (St. Louis, MO), dithiothreitol, iodoacetamide, sodium acetate, 2-morpholinoethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid and deuterium oxide from FUJIFILM Wako Pure Chemical Co., Ltd. (Tokyo, Japan), and MicroSpin G-25 columns from GE Healthcare (Chicago, IL). We purchased trypsin from Promega Co. (Madison, WI); angiotensin II from Peptide Institute, Inc. (Osaka, Japan); and 18O-water, acetonitrile (LC-MS grade), and ordinary water (LC-MS grade) from Merck (Darmstadt, HE).
7
Recombinant Protein Expression and Purification
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A single colony from verified BL21 (DE3) expression vector clones was cultured at 37°C overnight in 2 ml LB broth with 100 μg/ml carbenicillin. Then 30 μl of the overnight culture was inoculated to 3 ml fresh LB broth with 100 μg/ml carbenicillin. After ~4h of incubation at 37°C (OD~0.6), the culture was induced for protein expression with 100 μM IPTG and incubated for an additional 2 h. Bacteria were then spun down at 2,800 rpm for 10 min and the supernatant discarded. E. coli were lysed with of 0.3 μg/ml lysozyme in Tris-HCl buffer, pH 7.5 (Thermo Scientific), then proteins were heat-denatured at 100°C for 10 minutes and evaluated on SDS-PAGE gels by Coomassie staining. Specificity of protein expression was confirmed by western blot using mouse anti-His monoclonal antibodies (Sigma, St. Louis, MO), and rabbit anti-mouse IgG conjugated to horseradish peroxidase (HRP) (Sigma) as secondary antibodies; as well as rabbit anti-Listeriolysin polyclonal antibodies (Abcam, Cambridge, MA, USA), and goat anti-rabbit IgG-HRP (Millipore, Billerica, MA) as secondary antibodies. His-tagged Annexin 3 (A3) protein (Abcam) was used as positive control for antibody detection.
8
Alkali Lignin Extraction and Analysis
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Alkali Lignin was acquired from Sigma-Aldrich (St. Louis, MO, USA), potato dextrose agar (Difco, TX, USA), Hexamethylenetetramine (Fischer Scientific, Pittsburgh, PA, USA), Sodium hypochlorite, and sucrose were also purchased (Difco, TX, USA). The analytical grade of all the chemicals allowed for their use without any further purification, including sodium phosphate buffer (Sigma-Aldrich, St. Louis, MO, USA), Absolute ethanol (Riedel-de Häen, Niedersachsen, Germany), Tris-HCl buffer (Fischer Scientific, Pittsburgh, PA, USA), bovine serum albumin (BSA) (Sigma-Aldrich St. Louis, MO, USA), Formalin solution (neutral buffered, 10%, Merck, Boston, MA, USA), Whatman® qualitative filter paper, Grade 1 (Merck, Boston, MA, USA), and Ridomil Gold SL (2%) (45.3% mefenoxam, SYNGENTA, Wilmington, DE, USA). The Abcam H&E Staining Kit (Hematoxylin and Eosin) (Fischer Scientific, Pittsburgh, PA, USA was used, and the Milli-Q Plus system was used to create deionized water (Millipore, Milford, MA, USA).
9
Molecular Diagnostics Assay Development
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T4 DNA ligase, 10× ligation buffer, bovine serum albumin (BSA), ATP, GeneRuler low range DNA ladder, SYBR Gold gel stain, Tris-HCl buffer (1 M, pH 8.0), and Tris-acetate-EDTA buffer (TAE, 50×) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Bst 3.0 polymerase, Bst buffer (isothermal amplification buffer II, 10×), MgSO 4 , Nb.BtsI nickase, and loading buffer were obtained from New England BioLabs (Ipswich, MA, USA). Fetal bovine serum (FBS) and agarose were obtained from Sigma-Aldrich (St. Louis, MO, USA). Streptavidin-coated cross-linked hydroxyethyl starch iron oxide composite particles (100 nm size MNP, product code 10-19-102) were supplied by Micromod Partikeltechnologie GmbH (Rostock, Germany). Sequences of target, PLP, and detection probes listed in Table S1 were synthesized by Integrated DNA Technologies (Coralville, IA, USA) and dissolved in 50 mM Tris-HCl (pH 8.0).
10
Peptide Purification from A. ebracteatus Hydrolysate
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HPLC analysis with a UV detector (RP-HPLC) was performed to partially purified peptides from A. ebracteatus protein hydrolysate. The column employed was an ODS-2 HYPERSIL C18; 250×4.6 mm, 5 µm particle size (Thermo Fisher Scientific, Inc.). A gradient mobile phase of 0–50 min was applied, with HPLC-grade 0.1% trifluoroacetic acid (TFA) (Thermo Fisher Scientific, Inc.), acetonitrile (Thermo Fisher Scientific, Inc.) and water (90:10 v/v). The injected sample volume was 100 µl (17.05 µg protein/ml) and the flow rate was 1.00 ml/min. Detection was performed at 220 nm. Each fraction was collected using a fraction collector (Amersham Pharmacia Biotech). Finally, fractions were lyzed in 50 mM Tris/HCl buffer (Thermo Fisher Scientific, Inc.) and then lyophilized using a freeze dryer (LaboGene).
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