Anti-HLA antibodies (SN230G6, SN607D8 and W6/32) were spotted, via random coupling, using a Continuous Flow Microspotter (Wasatch Microfluidics) onto a SensEye Easy2Spot G-type sensor (Senss). Spotting was done in 10 mM acetate buffer phosphate-buffered saline (PBS) + 0.075%-Tween 80 (Amresco), pH 4.5 in duplicate at a two-fold dilution ranging from 60 nM to 7.5 nM for 15 min. The sensor was deactivated with 100 mM ethanolamine (Merck), pH 8.8 for 7 min. Binding measurements were carried out in an IBIS MX96 (IBIS Technologies). First, 100 nM HLA-A*02:01 was flowed over the sensor for 5 min, followed by a 5-min flow of 100 nM of each anti-HLA antibody separately. Regeneration was then performed with 10 mM Gly-HCl, pH 2.4. This was repeated for every possible antibody combination.
Tween 80
Manufactured by Avantor
Sourced in United States, United Kingdom, Germany, France, China, Italy, Bangladesh, Mexico
Tween 80 is a non-ionic surfactant. It is used as an emulsifier, solubilizer, and stabilizer in various pharmaceutical and cosmetic formulations.
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Lab products found in correlation
Glycerol, by Merck Group (7 mentions)
Methanol, by Merck Group (6 mentions)
Tween 20, by Avantor (5 mentions)
Middlebrook 7h9 medium, by BD (5 mentions)
Glycerol, by Avantor (4 mentions)
Sodium hydroxide (naoh), by Merck Group (4 mentions)
Methanol, by Avantor (4 mentions)
Acetic acid, by Merck Group (4 mentions)
Pentobarbital sodium, by Merck Group (3 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (3 mentions)
Formalin solution, by Merck Group (3 mentions)
Middlebrook 7h9 broth, by BD (3 mentions)
Dimethyl sulfoxide (dmso), by Avantor (3 mentions)
Hydrochloric acid (hcl), by Merck Group (3 mentions)
Bovine serum albumin (bsa), by Avantor (2 mentions)
Trehalose, by Avantor (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Glycerol, by Maclin Biochemical Technology (2 mentions)
74 protocols using tween 80
1
Quantifying Anti-HLA Antibody Binding
2
Nociceptive Behavior Evaluation Protocol
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One day before experiments, animals (2–4 months old) were transferred to the experimental room for acclimatization, separated in individual cages and provided food and water ad libitum. Experiments were performed during the day (light cycle). Stock solutions of CIM0216 (100 mM) alone or in mixture with Baclofen (25 mM) were diluted 20x in a sterile vehicle solution of 10% PEG-200 (Sigma-Aldrich), 2% Tween-80 (Amresco) in 0.9% NaCl. Hind paw subcutaneous dorsal injection of 10 μL was performed using a 30G needle coupled to a Hamilton syringe, and duration and numbers of nocifensive behavior (licking and lifting of injected paw) were recorded for a period of 10 min by an observer blind to the tested substances. Every animal was subject to random order injections of CIM0216 or its mixture with Baclofen in one paw followed 7 days later by the other reagent in opposite paw. A similar procedure was used for AITC and baclofen. We used both male and female animals and the data were pooled, as there was no significant difference between the two sexes. The final injected dose was 50 nmol/paw for CIM, 100 nmol/paw for AITC and 12.5 nmol/paw for Baclofen. All animal experiments were performed in accordance with the requirements of the Institutional Animal Care and Use Committee at Rutgers New Jersey Medical School
3
Formulation and Characterization of Anti-TB Drug Delivery
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The following reagents and materials were used: bovine serum albumin (BSA) (Amresco, Solon, OH, USA); polylactic acid (PLA, MW20000, Shandong Institute of Medical Instrument, China); INH, RFP (Wuhan Fuchi Biotechnology Co., Ltd., China); Tween 80 (Amresco); INH and RFP standard preparation (National Institutes for Food and Drug Control, approval No. 100578-200401; 110757-200206); bag filter (MD77,8000-14000, Beijing Solarbio Science and Technology Co., Ltd., China); trehalose (Amresco); mannitol (Shanghai Kaiyang Biotechnology Co., Ltd., China); and chromatographically pure methanol and methyl cyanide. Other reagents were analytically pure.
4
Immunoassay Reagents and Kits
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DBP (>99%), FITC, formalin solution (10%) and pentobarbital sodium were purchased from Sigma–Aldrich (St. Louis, MO, USA). Tween-80 was obtained from Amresco (Solon, OH, USA). All other chemicals were of analytical grade. Mouse enzyme-linked immunosorbent assay (ELISA) kits for total IgE was purchased from Biolegend (San Diego, CA, USA). Mouse ELISA kits for IL-4, interferon (IFN)-γ, IL-17A, TSLP and TNF-α were from eBioscience (San Diego, CA, USA). Mouse anti-IL-5-antibody, mouse anti-IL-13-antibody, and mouse anti-eosinophil cationic protein (ECP)-antibody, goat-anti-rabbit lgG-antibody, a rabbit lgG peroxidase conjugated streptavidin-biotin complex (SABC-POD) kit and a diaminobenzidine (DAB) kit were obtained from Boster Bio-engineering (Wuhan, China).
5
Mycobacterial Cultivation and Macrophage Infection
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M. abscessus ATCC19977, M. tuberculosis H37Rv ATCC27294, M. tuberculosis H37Ra ATCC25177, M. bovis ATCC19210, M. bovis BCG ATCC35737 (M. bovis BCG), and M. smegmatis mc2155 ATCC700044 were propagated in Middlebrook 7H9 broth (BD, New York, NJ, USA) supplemented with 10% OADC (BD, New York, NJ, USA), 0.2% glycerol (Sigma, Saint Louis, MO, USA), and 0.05% Tween 80 (Amresco, Houston, TX, USA) or on 7H11 agar plates supplemented with 0.5% glycerol and 10% OADC. THP-1 macrophages (ATCC TIB-202) were cultured in RPMI-1640 medium (Gibco, Shanghai, China) supplemented with 10% fetal bovine serum (Gibco, Waltham, MA, USA) at 37 °C with 5% CO2.
6
Sediment Dilution and Coliform Plating
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The sediment samples were diluted at 1:10 (1 g sediment + 9 mL Ringer Tween solution 0.3%) in 50 mL sterile plastic tubes (Greiner®, bio-One™, Kremsmünster, Austria) with 0.3% Ringer Tween (TWEEN 80®, Amresco, VWR™, Vienna, Austria) solution and incubated at the current river temperatures for one hour on a roll mixer (CATRM5®, servoLAB™, Kumberg, Austria). Approximately 500 µL of diluted samples was consequently plated onto Chromo Cult Coliform Agar plates. Post incubation, the colonies were isolated and stored using an identical procedure employed for the water samples.
7
Mouse Allergy Biomarkers Measurement
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DINP (> 99%), DBP (> 99%), FITC, pentobarbital sodium and formalin solution (4%) were bought from Sigma-Aldrich (St. Louis, MO, USA). Tween-80 and Tween-20 were purchased from Amresco (Solon, OH, USA). Mouse enzyme-linked immunosorbent assay (ELISA) kits for total IgE, IL-4 and (IFN)-γ were obtained from eBioscience (San Diego, CA, USA). MDA and GSH test kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The protein test kits were provided by Beijing Dingguo Changsheng Biotechnology Co. LTD (Beijing, China).
8
Standardized Antimicrobial Susceptibility Testing
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DST was performed by using broth micro-dilution methods according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines (Krishnan et al., 2009 (link); Schon et al., 2020 (link)). The strains were grown in Middlebrook 7H9 broth medium (Difco, Becton, Dickinson and Company, New Jersey, USA) supplemented with 10% OADC, (Difco), 0.2% glycerol and 0.05% Tween-80 (Amresco, USA) for initial culture. Later, Middlebrook 7H9 (final pH 6.6 ± 0.2) broth medium supplemented with 10% OADC, 0.2% glycerol without Tween-80 was used in the preparation of inocula for DST. Based on EUCAST guidelines the inocula were fixed up to OD600 0.6 with a final concentration of 1 × 105 to 5 × 105 CFU/mL. Cells in 96 well plates were incubated at 37°C. Several anti-microbials including rifabutin, imipenem, rifampicin (RIF), vancomycin, streptomycin, amikacin, clarithromycin, ethambutol, isoniazid, sulfamethoxazole, linezolid, and clofazimine were used to perform DST (Table S11
9
Preparation of Lipid-based Formulations
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Tween-80 (Amresco, #0442, USA) and Tween-20 (Amresco, #0777) were purchased. Lecithin from egg yolk was provided by Sinopharm Chemical Reagent (#6901933, Shanghai, China). Sesame oil was obtained from Blessing Mill (Wuhan, China). DMEM (#10569-044) and FBS (#12484-010) were purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). MTT (#M2128), dimethyl sulfoxide (DMSO, #D5879) of analytical reagent grade and Glycerol ReagentPlus (GC grade, #G7757) were obtained from Sigma–Aldrich (St Louis, MO, USA).
10
Murine Mycobacterium smegmatis Culture Protocol
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The mice used in this study were purchased from Vital River, Beijing, China. All experimental work involving animals was performed according to the guidelines recommended by the animal welfare and ethics of the Heilongjiang Animal Ethics Committee at the Heilongjiang science and technology government agency (Harbin, People’s Republic of China) and was approved and supervised by the commissioner for animal welfare at the Harbin Veterinary Research Institute (HVRI) representing the Institutional Animal Care and Use Committee. All the experiments were designed to minimize the numbers of animals used and every effort was made to minimize both pain and distress to the animals.
M. smegmatis cultures were grown in Middlebrook 7H9 medium (BD Biosciences) and supplemented with 10% oleic acid/albumin/catalase enrichment (10% OADC, BD Biosciences), 0.05% Tween 80 (Amresco), and 0.2% glycerol (Sigma-Aldrich). E. coli DH5α strain (Novagen) was used for plasmid preparations, and E. coli Rosetta 2 strain (Novagen) was used for protein expression. All E. coli strains were cultured in Luria-Bertani (LB) medium.
M. smegmatis cultures were grown in Middlebrook 7H9 medium (BD Biosciences) and supplemented with 10% oleic acid/albumin/catalase enrichment (10% OADC, BD Biosciences), 0.05% Tween 80 (Amresco), and 0.2% glycerol (Sigma-Aldrich). E. coli DH5α strain (Novagen) was used for plasmid preparations, and E. coli Rosetta 2 strain (Novagen) was used for protein expression. All E. coli strains were cultured in Luria-Bertani (LB) medium.
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