RPMI 1640 cell culture medium, 2-mercaptoethanol (ME), sodium pyruvate, and phosphate-buffered saline (PBS) were purchased from Thermo Fisher Scientific. Fetal calf serum (FCS), D-glucose, penicillin/streptomycin (P/S), phorbol 12-myristate 13-acetate (PMA), lipopolysaccharides (LPS) from E. coli O111:B4, Accutase, agarose, β-nicotinamide adenine dinucleotide sodium salt (NAD), lithium L-lactate, phenazine methosulfate (PMS), iodonitrotetrazolium chloride (INT), and bovine serum albumin (BSA) were purchased from Sigma-Aldrich/Merck. Tris base was ordered from Carl Roth (Germany). The TiO2 nanoparticles, carbon nanotubes, and MNP samples used in the following experiments are listed in Table 1 .
Tris base
Manufactured by Carl Roth
Sourced in Germany
TRIS base is a chemical compound that serves as a buffering agent in various laboratory applications. It is a white crystalline solid that is highly soluble in water and has a pH-stabilizing effect in solutions.
Automatically generated - may contain errors
Lab products found in correlation
Trichloroacetic acid, by Carl Roth (4 mentions)
Acetic acid, by Avantor (4 mentions)
Acetic acid, by Merck Group (4 mentions)
Genios plus microplate reader, by Tecan (4 mentions)
Iodonitrotetrazolium chloride, by Merck Group (1 mentions)
Phenazine methosulfate, by Merck Group (1 mentions)
Agarose, by Merck Group (1 mentions)
β nicotinamide adenine dinucleotide sodium salt, by Merck Group (1 mentions)
Lithium l lactate, by Merck Group (1 mentions)
Penicillin streptomycin p s, by Merck Group (1 mentions)
Fetal calf serum (fcs), by Merck Group (1 mentions)
Accutase, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Chloramphenicol, by Carl Roth (1 mentions)
Naclo4 h2o, by Merck Group (1 mentions)
4 2 pyridylazo resorcinol, by Merck Group (1 mentions)
1 4 dithiothreitol dtt, by Carl Roth (1 mentions)
Chelex 100 resin, by Bio-Rad (1 mentions)
7 protocols using tris base
1
Evaluating Nanomaterial Cytotoxicity in Cells
2
Metalloprotein Purification and Characterization
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Acetonitrile (MeCN), HCl (trace metal grade), NaClO4 H2O, ZnSO4·7H2O, 4-(2-pyridylazo)resorcinol (PAR), and Triton X-100 were from Merck Millipore. Tris(2-carboxyethyl)phosphine hydrochloride (TCEP), tryptone, yeast extract, LB Broth, agar, agarose, isopropyl-β-D-1-thiogalactopyranoside (IPTG) and sodium dodecyl sulfate (SDS) were purchased from Lab Empire. Ampicillin, chloramphenicol, 1,4-dithiothreitol (DTT) and Tris base were from Roth, pTYB21 vector and chitin resin from New England BioLabs, Chelex 100 resin from Bio-Rad, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid sodium salt (HEPES), 5,5ʹ-dithiobis-(2-nitrobenzoic acid) (DTNB) from TCI Europe. To eliminate trace metal ion contamination all pH buffers were treated with Chelex 100 resin (Bio-Rad) and degassed over 2 h prior to use. For the culture of E. coli, Luria–Bertani (LB) medium and agar plates were used.
3
Optical Fiber-Based CRP Biosensor
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Single-mode optical fibers (SMF-28e) were purchased from Corning Optical Communications (Berlin, Germany). Ethanol, glycerol, (3-glycidyloxypropyl)trimethoxysilane (GOPTS), L-cysteine, phosphate buffered saline (PBS; pH = 7.4), H2O2 (ω = 30%), potassium hydroxide, and urea were purchased from Sigma-Aldrich (Taufkirchen, Germany). Acetic acid, CaCl2·2H2O, 1 M HCl, MgCl2·6H2O, L-ascorbic acid, KCl, NaCl, 1 M NaOH, H2SO4 (ω = 96%) and Tris base were purchased from Carl-Roth (Karlsruhe, Germany). Hydrofluoric acid (HF; ω = 40%) and immersion oil were purchased from AppliChem (Darmstadt, Germany). Water was purified with a Milli-Q purification system. Human recombinant C-reactive protein (CRP; β = 1 mg/mL) was purchased from BioCat (Heidelberg, Germany) and pooled human >97% CRP deficient plasma was purchased from Dunn Labortechnik (Asbach, Germany). The CRP-specific single-stranded DNA aptamer (CRP-40-17-3′SH) with a thiol group at the 3′-end was synthesized and HPLC was purified by Metabion (Planegg, Germany) and delivered at 100 µM in bidest. water. The sequence of CRP-40-17-3′SH was: 5′-CCC CCG CGG GTC GGC TTG CCG TTC CGT TCG GCG CTT CCC CTT TTT TTT T-C6-SH-3′ [50 (link)].
4
Sulforhodamine B Viability Assay for NET/NEC Cells
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To assess tumor cell viabilities at 72 and 96 h post infection (hpi), the Sulforhodamine B (SRB) assay was employed. This viability assay measures cell density compared to mock treatment by quantifying the number of adherent (viable) cells [34 (link)]. For this purpose, NET/NEC cells were seeded in 24-well plates and infected with OV, mock treated, treated with everolimus or OV and everolimus together. At the respective time point of analysis (at 72/96 hpi), cells were fixed with 10% (v/v) trichloroacetic acid (Carl Roth, Karlsruhe, Germany) after rinsing them with 4 °C cold PBS. Fixation was allowed for at least 30 min at 4 °C. Next, cell cultures were washed with water. Then, fixed cells were stained with SRB dye (0.4% (w/v) in 1% (v/v) acetic acid; Sigma-Aldrich) for at least 10 min and rinsed afterwards with 1% (v/v) acetic acid (VWR, Radnor, PA, USA) to remove unbound SRB dye. After drying for another 24 h, 10 mM TRIS base (pH 10.5; Carl Roth) was added to solve remaining SRB dye. To measure the amount of bound SRB dye, the absorbance of the inoculum at a wavelength of 550 nm was determined in duplicates (using a Tecan Genios Plus Microplate Reader). As the SRB dye binds to cellular proteins, the absorbance correlates with cell density. In the figures, cell density of mock treated cells was adjusted as 100%; percentages refer to mock treatment.
5
Sulfo rhodamine B Viability Assay
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To assess tumor cell viabilities at 72 and 96 hours post infection (hpi), the Sulfo rhodamine B (SRB) assay was employed. This viability assay measures cell density compared to mock treatment by quantifying the number of adherent (viable) cells [34] . For this purpose, NET/NEC cells were seeded in 24well plates and infected with OV, mock treated, treated with everolimus or OV and everolimus together. At the respective time point of analysis (at 72/96 hpi), cells were xed with 10 % (v/v) trichloroacetic acid (Carl Roth, Karlsruhe, Germany) after rinsing them with 4 °C cold PBS. Fixation was allowed for at least 30 min at 4 °C. Next, cell cultures were washed with water. Then, xed cells were stained with SRB dye (0.4 % (w/v) in 1 % (v/v) acetic acid; Sigma-Aldrich) for at least 10 min and rinsed afterwards with 1 % (v/v) acetic acid (VWR, Radnor, PA, USA) to remove unbound SRB dye. After drying for another 24 hours, 10 mM TRIS base (pH 10.5; Carl Roth) was added to solve remaining SRB dye. To measure the amount of bound SRB dye, the absorbance of the inoculum at a wavelength of 550 nm was determined in duplicates (using a Tecan Genios Plus Microplate Reader). As the SRB dye binds to cellular proteins, the absorbance correlates with cell density. In the gures, cell density of mock treated cells was adjusted as 100 %;
percentages refer to mock treatment.
percentages refer to mock treatment.
6
Sulfo rhodamine B Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess tumor cell viabilities at 72 and 96 hours post infection (hpi), the Sulfo rhodamine B (SRB) assay was employed. This viability assay measures cell density compared to mock treatment by quantifying the number of adherent (viable) cells [34] . For this purpose, NET/NEC cells were seeded in 24well plates and infected with OV, mock treated, treated with everolimus or OV and everolimus together. At the respective time point of analysis (at 72/96 hpi), cells were xed with 10 % (v/v) trichloroacetic acid (Carl Roth, Karlsruhe, Germany) after rinsing them with 4 °C cold PBS. Fixation was allowed for at least 30 min at 4 °C. Next, cell cultures were washed with water. Then, xed cells were stained with SRB dye (0.4 % (w/v) in 1 % (v/v) acetic acid; Sigma-Aldrich) for at least 10 min and rinsed afterwards with 1 % (v/v) acetic acid (VWR, Radnor, PA, USA) to remove unbound SRB dye. After drying for another 24 hours, 10 mM TRIS base (pH 10.5; Carl Roth) was added to solve remaining SRB dye. To measure the amount of bound SRB dye, the absorbance of the inoculum at a wavelength of 550 nm was determined in duplicates (using a Tecan Genios Plus Microplate Reader). As the SRB dye binds to cellular proteins, the absorbance correlates with cell density. In the gures, cell density of mock treated cells was adjusted as 100 %;
percentages refer to mock treatment.
percentages refer to mock treatment.
7
Sulfo rhodamine B Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess tumor cell viabilities at 72 and 96 hours post infection (hpi), the Sulfo rhodamine B (SRB) assay was employed. This viability assay measures cell density compared to mock treatment by quantifying the number of adherent (viable) cells [34] . For this purpose, NET/NEC cells were seeded in 24well plates and infected with OV, mock treated, treated with everolimus or OV and everolimus together. At the respective time point of analysis (at 72/96 hpi), cells were xed with 10 % (v/v) trichloroacetic acid (Carl Roth, Karlsruhe, Germany) after rinsing them with 4 °C cold PBS. Fixation was allowed for at least 30 min at 4 °C. Next, cell cultures were washed with water. Then, xed cells were stained with SRB dye (0.4 % (w/v) in 1 % (v/v) acetic acid; Sigma-Aldrich) for at least 10 min and rinsed afterwards with 1 % (v/v) acetic acid (VWR, Radnor, PA, USA) to remove unbound SRB dye. After drying for another 24 hours, 10 mM TRIS base (pH 10.5; Carl Roth) was added to solve remaining SRB dye. To measure the amount of bound SRB dye, the absorbance of the inoculum at a wavelength of 550 nm was determined in duplicates (using a Tecan Genios Plus Microplate Reader). As the SRB dye binds to cellular proteins, the absorbance correlates with cell density. In the gures, cell density of mock treated cells was adjusted as 100 %;
percentages refer to mock treatment.
percentages refer to mock treatment.
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