A freshly prepared stock solution of CR (7.0 mg/mL) in 10 mM phosphate buffer at pH 7.4 was filtered through 0.22 μm syringe and then diluted in water at 5.0 μM final concentration. This solution was incubated for 30 minutes at room temperature with 0.5 mg/mL of Aβ16‐21 peptide. After incubation, UV‐Vis spectra of Congo Red (CR) alone or incubated with Aβ16‐21 were recorded between 400 and 750 nm on Nanodrop 2000c spectrophotometer equipped with a 1.0 cm quartz cuvette (Hellma).
Nanodrop 2000c spectrophotometer
Manufactured by Hellma
Sourced in United States
The Nanodrop 2000c spectrophotometer is a compact and easy-to-use instrument designed for the quantification and analysis of nucleic acids, proteins, and other biomolecules. It utilizes a unique patented sample retention system that requires only 1-2 microliters of sample volume to perform measurements. The Nanodrop 2000c provides accurate and reproducible results, making it a popular choice for applications in molecular biology, biochemistry, and life science research.
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Lab products found in correlation
7 protocols using nanodrop 2000c spectrophotometer
1
Spectrophotometric Analysis of Amyloid-β
2
Synthesis of Radiolabeled Peptide Conjugates
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Protected Nα-Fmoc-amino acid derivatives, coupling reagents, and Rink amide 4-methylbenzhydrylamine (MBHA) resin were purchased from Calbiochem-Novabiochem (Laufelfingen, Switzerland). The Fmoc-21-amino-4,7,10,13,16,19-hexaoxaheneicosanoic acid (Fmoc-Ahoh-OH) and Fmoc-8-amino-3,6-dioxaoctanoic acid (Fmoc-AdOO-OH) were purchased from Neosystem (Strasbourg, France). DOTA(OtBu)3-OH (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate tert-butyl ester) was purchased from CheMatech (Dijon, France). Citrate acid, sodium citrate, and sodium chloride were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). All other chemicals were commercially available by Sigma-Aldrich, Fluka (Bucks, Switzerland) or LabScan (Stillorgan, Dublin, Ireland) and were used as received unless otherwise stated. Preparative HPLCs were carried out on a LC8 Shimadzu HPLC system (Shimadzu Corporation, Kyoto, Japan) equipped with a UV Lambda-Max Model 481 detector. UV-Vis measurements were carried out on Thermo Fisher Scientific Inc. (Wilmington, Delaware USA) Nanodrop 2000c spectrophotometer equipped with a 1.0-cm quartz cuvette (Hellma). 177LuCl3 and 111InCl3 were obtained from IDB (Petten, The Netherlands) and Mallinckrodt Radiopharmaceuticals Italia (Segrate, Italy), respectively.
3
Peptide Concentration Determination
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PEG6-Y4 solutions were prepared dissolving the pure peptide powder in water at the desired concentration. The effective peptide concentration in solution was spectroscopically determined by UV–Vis measurements on Thermo Fisher Scientific Inc (Wilmington, Delaware USA) Nanodrop 2000c spectrophotometer equipped with a 1.0 cm quartz cuvette (Hellma) using as molar absorptivity (ε) the value of 5360 M−1·cm−142 (link).
4
Peptide Characterization and Preparation
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PEG24‐F6 solution was prepared by dissolving the lyophilized powder in double distilled water sonicating the sample for 1 minutes. Then peptide concentration was determined with UV‐Vis Thermo Fisher Scientific Inc (Wilmington, Delaware USA) Nanodrop 2000c spectrophotometer equipped with a 1.0 cm quartz cuvette (Hellma) using a molar absorptivity (ϵ257) of 1170 M−1 cm−1. The peptide solution was diluted at 10 mg/mL. Solutions of H+‐Aβ16‐21‐O− at 5.0 and 100 mg/mL were prepared in water or in HFIP, respectively. The peptide concentration in water was assessed by UV‐Vis spectroscopy using of 390 M−1⋅cm−1 as molar absorptivity at 257 nm. Finally, stock solution of H+‐F6‐O− was prepared dissolving the peptide powder directly in 1,1,1,3,3,3‐hexafluoro‐2‐propanol (HFIP) at 50 mg/mL. Subsequently, this solution was also diluted with HFIP to obtain a concentration of 5.0 mg/mL. For solid state characterization, these peptide solutions were drop‐casted onto flat microscope glass slides and allowed to air dry at room temperature.
5
Quantification of Peptide Concentrations
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Peptide solutions were prepared by dissolving 10 mg of each lyophilized peptide in 1.0 mL of water. These suspensions were sonicated for 30 min at room temperature; after centrifugation (5 min at 13,000 rpm), the concentration was spectroscopically determined on UV–Vis Thermo Fisher Scientific Inc. (Wilmington, DE, USA) NanoDrop 2000 c spectrophotometer equipped with a 1.0 cm quartz cuvette (Hellma). The quantification of the concentration was assessed using the molar absorptivity (ε) of 7800 M−1 cm−1 for Fmoc group at λ = 301 nm.
6
Preparation and Characterization of Amyloid-beta Peptide Solutions
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Ac-F6-O−, H+-F6-Am and Ac-F6-Am solutions were prepared dissolving peptide powders directly in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) at 100 mg/mL. For H+-F6-O−, solution concentration was 50 mg/mL. Subsequently, these solutions were properly diluted with HFIP at the final required concentration. Ac-Aβ16–21-Am peptide solution was prepared directly dissolving lyophilized powder in double distilled water. The experimental concentration was assessed by UV–Vis spectroscopy carried out on UV–Vis Thermo Fisher Scientific Inc (Wilmington, Delaware USA) Nanodrop 2000c spectrophotometer using a 1.0 cm quartz cuvette (Hellma) and a molar absorptivity (ε257) of 390 M−1/cm. Then, these peptide solutions were drop-casted onto flat microscope glass slides and allowed to air dry at room temperature.
7
Quantifying Peptide Conjugate Concentration
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Peptide conjugate solutions were prepared by dissolving the lyophilized powder in double distilled water. The concentration of the final solution was estimated by absorbance on UV-vis Thermo Fisher Scientific Inc (Wilmington, DE, USA) Nanodrop 2000c spectrophotometer equipped with a 1.0 cm quartz cuvette (Hellma) using a molar absorptivity (ε276) of 4230 M−1cm−1.
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