Sample Preparation. 1. Add 1 µL Activator to 100 µL patient sample or control. Mix and allow to sit at room temperature for 10 minutes.
Sample Preparation a. Laboratory Molded Specimens - Use cylindrical specimens that have been compacted using the gyratory compactor (AASHTO T 312). Specimen diameter must be 6 inches (150 mm) and a specimen height must be 4.5 inches +/- 0.2 inches (115 +/- 5 mm). Note 1 - Experience has shown that molded laboratory specimens of a known density usually result in a greater density (or lower air voids) after being trimmed. Therefore, it is recommended that the laboratory technician produce molded specimens with an air void level slightly higher than the targeted trimmed specimen. Determine the density of the final trimmed specimen in accordance with AASHTO T 166.
Sample Preparation. (1) Sample to be processed should correlate with stage and site to be treated.
Sample Preparation. Users are responsible for preparing their own samples; however, the Facility Manager must approve the prepared samples (see #6) and may disallow samples that are not stable and could damage the instrument. The Facility Manager will train Members in sample preparation and may provide assistance in special cases, but in general Members are responsible for their own samples.
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Sample Preparation. Human calmodulin (CaM) was expressed and purified in Escherichia coli BL21(DE3) cells, as described F xxxxx://xxx.xxx/10.1021/jacs.2c02201 previously.9 Two site-directed mutations at A17C and A128C in the NTD and CTD domains of CaM, respectively, were introduced as sites of attachment for the nitroxide spin-labels.9 Full deuteration was achieved by growing the bacteria in deuterated minimal medium, with U-[12C/2H]-glucose as the sole carbon source. R1 nitroxide labeling was carried out with S-(1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H- pyrrol-3-yl)methyl methanesulfonothioate (MTSL; Toronto Research Chemicals), as described previously.9 The purity of A17C-R1/ A128C-R1 nitroxide-labeled calmodulin was verified by mass spectrometry. Solutions for DEER EPR comprised 50 μM CaM (A17C-R1/ A128-R1), 8 mM CaCl2, 100 mM NaCl, 25 mM d-HEPES pH 6.4, and 20% (v/v) d8-glycerol, placed in 1.0 mm inner diameter (1.2 mm outer diameter) quarts EPR tubes (VitoCom). Freezing of the DEER samples was carried out using three different approaches: slow rate freezing (∼40 s) by placing the EPR tube in a −80 °C freezer;43 intermediate rate freezing (∼1.5 s) by directly placing the EPR tube in liquid N2;43 and rapid freeze-quenching (∼0.5 ms) by spraying a high- speed jet onto a spinning copper plate cooled to 77 K, as described previously.18,45 Q-Band DEER. Pulsed EPR data were collected at Q-band (33.8 GHz) and 50 K on a Bruker E-580 spectrometer equipped with a 150 W traveling-wave tube amplifier, a model ER5107D2 resonator, and a cryofree cooling unit, as described previously.52 DEER experiments were acquired using a conventional four-pulse sequence (Figure S1).53 The observer and XXXXX pump pulses were separated by ca. 90 MHz, with the observer π/2 and π pulses set to 12 and 24 ns, respectively, and the XXXXX π pulse to 10 ns. The pump frequency was centered at the Q-band nitroxide spectrum located at +40 MHz from the center of the resonator frequency. The τ1 value of 350 ns for the first echo period time was incremented eight times in 16 ns steps to average 2H modulation; the position of the XXXXX pump pulse was incremented in steps of Δt = 10 ns. The bandwidth of the overcoupled resonator was 120 MHz. All DEER echo curves were acquired for tmax = 7.5 μs, with the exception of the DEER echo curve for τ2 < 7.5 μs, where tmax was set to the value of τ2. DEER data were recorded with values of the dipolar evolution time T (=2τ2) set to 10, 15, 20, 25, 30, and 35 μs. Measurement t...
Sample Preparation. 1. Prepare hemolysates of patient specimens and controls as instructed in the "Specimen Preparation" section.
Sample Preparation. ≥ Anthracene (Ac) (Aldrich, scintillation grade, purity 99.0%) was purified in a home-built zone-refiner for about 4000 passes. 7.8,15.16-dibenzoterrylene (DBT) was purchased from Xx. X. Schmidt (Laboratory for PAH Research, Greifenberg, Germany). Samples were single crystals of Ac as a host, doped with DBT. Flat crystals (flakes) were grown by co-sublimation in a home-built device under a 150 mbar nitrogen atmosphere. This co-sublimation device consists of a large tube (30 mm diameter) containing a load of Ac powder, heated by resistant wires. Crystals grow on a collector plate at a slightly lower temperature than the load [70]. Within this tube, a small crucible containing the guest, DBT, is heated with resistant wires wrapped around it, up to an adjustable temperature. We thus can control the concentration of probe molecules in the matrix. The sublimation flakes develop along the (a-b) plane and reach diameters of a few millimeters and thicknesses from a few micrometers to a few tens of micrometers. They are optically contacted to a glass substrate or glued to it by a small amount of vacuum grease, mounted and cooled down to 1.4 K in a helium bath cryostat.
Sample Preparation. Draw down the paint on four glass panels measuring approximately 100 by 200 mm to a dry film thickness of 0.102 mm to 0.107 mm. Downloaded from xxxx://xxx.xxxxxxxxx.xxx TT-P-1952F
Sample Preparation. Use the test panels prepared for the accelerated weathering test (4.3.10.1).
