The amount of 1 × 106 cpm of 32P-labeled oligonucleotides was dissolved in 1× PBS buffer containing 100 mM potassium chloride. The samples were denatured at 90°C for 6 min and then cooled to room temperature overnight. After this time, 200 μL of human serum from human male AB plasma (Sigma-Aldrich) was added, and the samples were incubated at 37°C. Aliquots of 5 μL were removed after 0, 10, 20, 40, 60, 120, 180, and 1,440 min of incubation and were then mixed with 5 μL of a 70% deionized formamide solution containing 50 mM EDTA and cooled on dry ice to quench the reaction. The samples were loaded on a 12% denaturing polyacrylamide gel prepared in 1× TBE buffer. PAGE in denaturing condition was performed in 1× TBE buffer at 20 W for 3 hr at room temperature. The resultant gel was imaged and quantified by storage phosphor technology using a Fuji PhosphorImager and MultiGauge Analysis Software.
Human serum from human male ab plasma
Manufactured by Merck Group
Sourced in United States
Human serum from human male AB plasma is a laboratory product derived from the blood of male individuals with AB blood type. It is a rich source of various proteins, enzymes, and other biomolecules found in the human circulatory system. This product can be used as a component in cell culture media, immunoassays, and other in vitro applications requiring human-derived biological materials.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Promega (2 mentions)
M csf, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Potassium chloride (kcl), by Merck Group (2 mentions)
Fuji phosphorimager, by Fujifilm (1 mentions)
Multigauge analysis software, by Fujifilm (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Cd14 microbead, by Miltenyi Biotec (1 mentions)
K4fe cn 6, by Merck Group (1 mentions)
Tris buffer saline, by Merck Group (1 mentions)
K3fe cn 6, by Merck Group (1 mentions)
2 5 dihydroxybenzoic acid dhb, by Merck Group (1 mentions)
Acetonitrile acn, by Merck Group (1 mentions)
Carbon nanoparticles, by Merck Group (1 mentions)
Ribonuclease b rnase b from bovine pancreas, by Merck Group (1 mentions)
Human alpha 1 acid glycoprotein agp, by Merck Group (1 mentions)
Pngase f with 10 g7 reaction buffer, by New England Biolabs (1 mentions)
Iodomethane, by Merck Group (1 mentions)
10 protocols using human serum from human male ab plasma
1
Stability of 32P-labeled Oligonucleotides in Human Serum
2
Differentiation of Monocyte-Derived Macrophages
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Monocyte-derived macrophages (MDMs) were differentiated from monocytes isolated from human peripheral blood mononuclear cells (PBMCs) using CD14 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany). The human PBMCs were collected from eight different healthy donors (Donors 1–8). The monocytes were differentiated by treatment with human macrophage colony-stimulating factor (M-CSF; PeproTech, London, UK) for 1 week. The MDMs were maintained in RPMI 1640 (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) (Promega, Madison, WI, USA), 5% human serum from human male AB plasma (Sigma Aldrich, St. Louis, MO, USA), 0.2 % GlutaMax (Thermo Fisher Scientific), and 10 ng/mL M-CSF. HeLa and 293T cells were maintained in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific) supplemented with 10% FBS (Promega). All cells used in this study were cultured at 37 °C in a humidified atmosphere of 5% CO2.
3
COVID-19 Spike Protein and Antibody Synthesis
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COVID-19 recombinant protein (COVID-19 Spike Protein RBD Domain Coronavirus) and monoclonal COVID-19 antibody (Humanized COVID-19 Spike S1 Protein Coronavirus Monoclonal Antibody) were purchased from MyBioSoruce (San Diego, USA). HAuCl4 and NaBH4 (Au–NPs precursors), KCl, K4Fe(CN)6, K3Fe(CN)6, glutaraldehyde (25%), cysteamine hydrochloride, bovine serum albumin (BSA), human serum from human male AB plasma (hemoglobin ≤ 20 mg·dL−1), phosphate buffer saline (PBS, pH 7.4) and tris-buffer saline (TBS) were supplied by Sigma-Aldrich (St. Louis, USA). DMF (99.5%) was acquired from Lach-Ner (Neratovice, Czech Republic). All the solutions were prepared in PBS pH 7.4 with the exception of BSA solution, which was prepared in TBS 1x.
4
Glycoprotein Analysis via Mass Spectrometry
5
SARS-CoV-2 Serological Assay Validation
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Three positive controls in the form of intravenous blood sera from confirmed SARS-CoV-2 PCR-positive patients (collected 10–30 days post onset of symptoms) were obtained from Naha Municipal Hospital, Naha, Japan. After titer analysis of all three samples (data not shown), the two samples with the strongest titers were pooled and used as the positive control for all assays. Positive controls (collected at least 90 days after onset of symptoms) for validation of the capillary blood collection method were obtained from Okinawa Chubu Hospital, Uruma City, Japan. Negative controls taken from patients prior to November 2019 were obtained from Naha Municipal Hospital from intravenous blood, and from a commercial serum pool (Human Serum from human male AB plasma, Sigma Aldrich H4522-100ML, Batch #SLCD1948, serum was pooled prior to August 2019). Human MERS-convalescent serum and SARS-CoV-2 convalescent plasma (NIBSC code 20/130) were obtained from the National Institute of Biological Standards and Control, UK.
6
Microfluidic Malaria Detection Assay
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The microfluidic microplate plate (Optimiser™ microplate), holder, and absorbent pad were provided from MiCo BioMed Co., Ltd. (Seoul, Korea). Mouse monoclonal PfLDH antibody was purchased from Fapon Biotech (BRCMALS212, Guangdong, China), and rabbit polyclonal PfLDH antibody was purchased from LifeSpan BioScience, Inc. (LS-C488775, Seattle, USA). PfLDH and PvLDH were provided by BioNano Health Guard Research Center (H-GUARD). Polyclonal anti-rabbit HRP-tagged antibody (HRP-Ab) was obtained from Cell Signaling Technology, Inc. (Danvers, USA). Phosphate-buffered saline (PBS), blocking solution (StartingBlock™) and HRP substrate kit (QuantaRed™ Enhanced Chemifluorescent HRP substrate kit) were purchased from Thermo Fisher Scientific Ltd. (Waltham, USA). Bovine serum albumin (BSA) and sodium carbonate were purchased from Sigma-Aldrich (Louis, USA). Human serum (from human male AB plasma, USA origin, sterile-filtered) was purchased from Sigma-Aldrich (Louis, USA).
7
HPLC-Purified DNA Aptamer Characterization
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The used HPLC-purified DNA aptamers were obtained from FRIZ Biochem (Neuried, Germany). The sequences were as follows:
S1: 5′-OH-(CH2)6-S-S-(CH2)6- CGA CTG GTA GGC AGA TAG GGG AAG CTG ATT CGA TGC GTG GGT CG -MB-3′
S2: 5′-OH-(CH2)6-S-S-(CH2)6- TAG GCA GAT AGG GGA AGC TGA TTC GAT GCG TG-MB-3′
S3: 5′-OH-(CH2)6-S-S-(CH2)6- GCA GAT AGG GGA AGC TGA TTC GAT GC-MB-3′
The aptamer concentration was obtained by recording the absorbance at a 260 nm wavelength with UV/vis spectroscopy (DS-11 Series Spectrophotometer/Fluorometer, DeNovix Inc., Wilmington, DE, USA). All tested solutions were prepared with Milli-Q water produced by a Milli-Q ultrapure water system (18.25 MΩ cm, Gradient A10, Merck Millipore, Burlington, MA, USA). Monofunctional methoxy-polyethylene glycol thiol (PEG, 2 kDa), tris-(2-carboxyethyl) phosphine hydrochloride (TCEP), 6-mercaptho-1-hexanol (MCH), potassium chloride, magnesium chloride, potassium ferrocyanide (K4[Fe(CN)6]), sodium chloride, potassium ferricyanide (K3[Fe(CN)6]), and human serum from human male AB plasma were obtained from Sigma-Aldrich Chemie GmbH (Darmstadt, Germany). Ethanol and isopropanol were ordered from Merck (Darmstadt, Germany).
S1: 5′-OH-(CH2)6-S-S-(CH2)6- CGA CTG GTA GGC AGA TAG GGG AAG CTG ATT CGA TGC GTG GGT CG -MB-3′
S2: 5′-OH-(CH2)6-S-S-(CH2)6- TAG GCA GAT AGG GGA AGC TGA TTC GAT GCG TG-MB-3′
S3: 5′-OH-(CH2)6-S-S-(CH2)6- GCA GAT AGG GGA AGC TGA TTC GAT GC-MB-3′
The aptamer concentration was obtained by recording the absorbance at a 260 nm wavelength with UV/vis spectroscopy (DS-11 Series Spectrophotometer/Fluorometer, DeNovix Inc., Wilmington, DE, USA). All tested solutions were prepared with Milli-Q water produced by a Milli-Q ultrapure water system (18.25 MΩ cm, Gradient A10, Merck Millipore, Burlington, MA, USA). Monofunctional methoxy-polyethylene glycol thiol (PEG, 2 kDa), tris-(2-carboxyethyl) phosphine hydrochloride (TCEP), 6-mercaptho-1-hexanol (MCH), potassium chloride, magnesium chloride, potassium ferrocyanide (K4[Fe(CN)6]), sodium chloride, potassium ferricyanide (K3[Fe(CN)6]), and human serum from human male AB plasma were obtained from Sigma-Aldrich Chemie GmbH (Darmstadt, Germany). Ethanol and isopropanol were ordered from Merck (Darmstadt, Germany).
8
Immortalized Erythroid Cell Line Differentiation
9
MHC II Allele-Optimized PBMC Isolation
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Tryptic soy broth (TSB) was purchased from Research Products International. BBL Mueller Hinton II Broth Cation-Adjusted (caMHIIB) was purchased from Becton, Dickinson and Company (BD). Bovine serum albumin (BSA) and human serum (from human male AB plasma, U.S. origin, sterile-filtered) were purchased from Sigma-Aldrich. Ninety-six–well plates with lids, sterile, Greiner Bio-One, were purchased from VWR International. Daptomycin was purchased from Selleck Chemicals. Goat anti-mouse horseradish peroxidase–conjugated IgG antibody was from Thermo Fisher Scientific. Interleukin-2 (IL-2) ELISpot (enzyme-linked immunospot) kits were from Mabtech, and IL-2 was from PeproTech. Human donor PBMCs were purchased from Cellular Technology Limited and were chosen to represent MHC II genotypes that both (i) included MHC II alleles for which the designs were explicitly optimized and (ii) excluded MHC II alleles for which the designs were explicitly optimized. The sample proportion and U.S. population proportion [taken from (51 (link))] of the selected donors’ DRB1 MHC II alleles are shown in fig. S2.
10
Tyrosinase-based Biosensor Fabrication
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Tyrosinase from mushroom (≥1000 unit/mg), catechol and captopril were purchased from Sigma-Aldrich (St. Louis, MO). Potassium hexachloroiridate-(IV), sodium hydrogencitrate, graphene oxide (4 mg/mL, dispersion in H2O), (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide) (EDC), N-hydroxysuccinimide (NHS), ascorbic acid, uric acid and paracetamol were also purchased from Sigma-Aldrich (St. Louis, MO). Milli-Q water was obtained from purification system and all solutions were prepared with ultra-pure water from a Millipore-Milli-Q system. Human serum from human male AB plasma was purchased from Sigma-Aldrich (therefore, no need for ethical committee permission, St. Louis, MO). The inks for screen printed electrode preparation were purchased from Acheson Industries, Germany.
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