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Separation Of The Bound And Unbound Forms Of The Radioactivity
Published in Erwin Regoeczi, Iodine-Labeled Plasma Proteins, 2019
A length of tubing is cut that can accommodate, between knots at both ends, the fluid to be placed in it in a space that is 2 to 3 times larger than the volume of the fluid. When a bag prepared this way is submerged in the outer compartment, the outer pressure compresses the bag and flattens its contents in a thin layer. Some extra length of tubing is required at both ends to keep the tubing in position in the measuring cylinder used as an outef compartment (Figure 36). Before use, the tubing is rolled up and soaked for approximately 10 min in the buffer to be used as the outer compartment. Dialysis of delicate proteins may require soaking in edetic acid (EDTA; 0.1 M), followed by washings in distilled H2O. If a tubing dries out, it should not be resoaked as it could leak. After the lower end has been tied, the tubing is filled with buffer and tested, by applying moderate finger pressure, for leakage. Care should be taken not to contaminate the upper segment of the tubing when applying the sample. The final arrangement, in which the bag is prevented from floating up to the surface, is illustrated in Figure 36. The spout of the cylinder should be sufficiently drained after each decanting so the diffusate does not trickle down on the outer wall.
Plasma and Blood Viscosity
Published in Gordon D. O. Lowe, Clinical Blood Rheology, 2019
The recommended anticoagulant for plasma viscosity measurement is edetic acid (EDTA, K, or Na salt) at a concentration of 3.4 to 4.8 mmol/ℓ blood (1.5 to 2 g/ℓ), since this causes least change in plasma viscosity.6,12 The dry salt is recommended to avoid dilution. Conveniently, this anticoagulant is also recommended for routine blood cell counts and for measurement of blood viscosity, since it has minimal effects on red cell morphology or rheology. Containers should be plastic or glass; small enough to leave little space above the sample; flat-bottomed to allow standing on the bench, reducing contact of plasma and sedimented cells; sufficiently strong to allow centrifugation; and with a close-fitting stopper to prevent leakage or evaporation.12
Discovery of novel ATAD2 bromodomain inhibitors that trigger apoptosis and autophagy in breast cells by structure-based virtual screening
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2020
Dahong Yao, Jin Zhang, Jinhui Wang, Dabo Pan, Zhendan He
Adherent and floating cells were collected after treatment with AM879 for indicated times. Cells were lysed in a lysis buffer consisting of Hepes 50 mM pH 7.4, Triton-X-100 1%, sodium orthovanada 2 mM, sodium fluoride 100 mM, edetic acid 1 mM, PMSF 1 mM, aprotinin (Sigma, MO, USA) 10 mg/L and leupeptin (Sigma) 10 mg/L at 4 °C for 1 h. After 12,000 rpm centrifugation for 15 min, the protein content of supernatant was determined by the Bio-Rad DC protein assay (Bio-Rad Laboratories, Hercules, CA, USA). Equal amounts of the total protein were separated by 10–15% SDS-PAGE and transferred to PVDF membranes, and the membranes were soaked in blocking buffer (5% skimmed milk or BSA). Proteins were detected using primary antibodies, followed by HRP-conjugated secondary antibody and visualised by using ECL as the HRP substrate. Quantity One 4.4 was used to quantify.