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34 protocols using dynapro titan

1

Synthesis of Nanosized Transition Metal Dichalcogenides

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Nanosized TMDs were prepared according to the procedure published in the literature.69 Briefly, raw solid MoS2 or WS2 was mixed with sodium cholate in 5 : 1 (w/w) ratio in 300 mL of water and sonicated for 20 h using the ultrasonic processor (120 W and 20 kHz with pulse-on for 2 s and pulse-off for 4 s) in the ice bath to prevent overheating. The resultant black dispersion was centrifuged at 3000 rpm for 30 min. The yellow-green supernatant containing dispersed TMDs was separated from the precipitate and subjected to another centrifugation using a higher speed at 12 000 rpm for 30 min. The nanosized TMD pellet at the bottom of the tube was redispersed in water and sonicated to extract the intercalated sodium cholate. This new dispersion was centrifuged at 12 000 rpm for 30 min and washed with water. The centrifugation and washing step was repeated at least three times to remove sodium cholate completely. Resulting precipitate was dispersed in 300 mL of deionized water to prepare a suspension of nanosized TMD particles. Hydrodynamic size of the nanosized TMDs was measured using dynamic light scattering (DLS), DynaPro Titan, Wyatt Technology Corporation, USA.
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2

Multi-Technique Nanomaterial Characterization

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Images were obtained using a JEM-2100F transmission electron microscope (TEM; JEOL Ltd., Tokyo, Japan). The Brunauer–Emmett–Teller surface area, N2 adsorption–desorption isotherms, and pore volume were obtained using the Micrometritics ASAP 2010 instrument (Micrometritics, Norcross, GA, USA). Fourier transform infrared (FT-IR) spectra were acquired using the Nicolet iS10 instrument (Thermo Fisher Scientific, Waltham, MA, USA). In addition, thermogravimetric analysis (TGA) was performed using the STA449C thermal analysis instrument (Netzsch, Germany). High-performance liquid chromatography (HPLC) was performed using the Agilent HP1100 instrument (Agilent Technologies, Santa Clara, CA, USA). Dynamic light scattering (DLS) was performed using the TC type Dynapro Titan instrument (Wyatt Technology, Santa Barbara, CA, USA).
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3

Hydrodynamic Radii of Recombinant Proteins

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Hydrodynamic radii of four proteins (RabGAP, RabGAP-CTR1, RabGAP-CTR2 and CTR) were estimated by using dynamic light scattering (DLS). The purified His6-cleaved recombinant proteins in GF buffer were analyzed using DynaPro Titan (Wyatt Technology Corporation) instrument.
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4

Dynamic Light Scattering of VNPs

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Dynamic Light Scattering (DLS) was measured using a DynaPro Titan, Wyatt Technology Corporation (laser wavelength 830 nm, scattering angle 20°) and Dynamics software. 13 μL samples of 0.2 mg mL–1 of VNPs were analysed at 25 °C. Data were presented as an average of three measurements of 10 runs each.30 (link)
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5

Glycomodule Conformational Changes

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The effect of temperature on glycomodule conformations was measured using a DynaPro Titan and ProteinSolutions Temperature controlled MicroSmapler (Wyatt Technology, Santa Barbara, CA). Five hundred µg of each glycomodule were dissolved in 1 ml of d.d. water. Radii of gyration were measured at 5°C, 8°C, 10°C, 14°C, 20°C, and 24°C. Five readings were taken at each temperature.
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6

Characterization of Fn and HccFn Nanocages

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The prepared native Fn and HccFn nanocages were characterized using transmission electron microscope (TEM), and dynamic light scattering (DLS).
TEM. For TEM observation, the native Fn and HccFn nanocage samples (20μl, 0.1mg/mL) were embedded in Plasma Cleaner HPDC32G treated copper grid and stained with 1% uranyl acetate for 1 min, then imaged with a JEM-1400 80-kV TEM (JEOL, Japan).
DLS. The native Fn and HccFn protein samples (100μl, 0.25 mg/mL) were prepared in PBS buffer. DynaPro Titan (Wyatt Technology) was used to perform DLS analysis. The temperature was controlled at 25°C.
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7

Dynamic Light Scattering Analysis of Protein Samples

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These measurements were performed using a DynaPro Titan (Wyatt Technology Corporation) with temperature control and a microsampler with a diode laser at a 90° scattering angle, a wavelength at 657 nm, and a power of 15 mW. The measurements were performed with samples (7–10 mM) dissolved in PBS at pH 7.4 in a temperature range of 25–75 °C. The samples were filtered with a 0.45 µm Nylon filter using Fisherbrand 10 mm o.d. glass tubes prior to the experiments. The obtained DLS measurements were obtained from Dynamics V6 software in the form of histograms. The graphs of average hydrodynamic diameters (DH) as a function of temperature in Figure 4 were constructed using Kaleida graph version 3.6.5, and the size distribution histograms of Figure 5 were constructed using PRISM version 4.0.
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8

Structural and Elemental Analysis of Ferritin

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TEM: the ferritin samples (10 μL, 0.1 mg mL-1) were embedded in a Plasma Cleaner HPDC32G-treated copper grid and stained with 1% uranyl acetate for 1 min, then imaged with a JEM-1400 80 kV TEM (JEOL, Japan). DLS: the average diameter of the ferritin samples (20 μL, 0.1 mg mL-1) was analyzed by the DynaPro Titan with a temperature-controlled micro-sampler of 25 °C (Wyatt Technology, U.S.A.). Inductively coupled plasma mass spectrometry (ICP-MS): the element contents of ferritin samples from different species were analyzed by the Agilent ICPMS7800.
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9

Dynamic Light Scattering for Particle Sizing

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Hydrodynamic radius (RH) measurements were made as previously described [44 (link)]. Measurements were taken at room temperature with a DynaPro Titan instrument (Wyatt Technology, Santa Barbara, CA). Samples (30 μl) were placed into a quartz cuvette and light scattering intensity was collected at a 90º angle using a 5-second acquisition time. Particle diffusion coefficients were calculated from auto-correlated light intensity data and converted to RH with the Stokes-Einstein equation using Dynamics software (version 6.12.0.3). Histograms of percent intensity vs. RH were generated by data regularization and intensity-weighted mean RH values were derived from the regularized histograms.
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10

Characterizing SELP Nanoparticle Size

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The average particle size and size distribution of the SELP and ANS-loaded SELP micellar-like nanoparticles were measured using a DynaPro Titan dynamic light scattering (DLS) instrument (Wyatt Technology, Santa Barbara, CA, USA). Samples were filtered using a 0.45 µm syringe filter prior to analysis and all measurements were completed at 25 °C. SELP and ANS-loaded SELP nanoparticles were analyzed at 0.1–1 mg/mL. Herein we report DLS data as an average of three measurements per sample type.
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