The molecular mass of EPOLs was decreased by treating their solutions (1 g/L) with a Branson sonifier equipped with a microtip at 2.8 Å. Samples were cooled in an ice bath and sonicated using 5 bursts of 1 min each, separated by 1 min intervals. They were subsequently exchanged three times with 99.9% D2O by lyophilisation and finally dissolved in 0.7 mL 99.96% D2O. Spectra were recorded on a 500 MHz VARIAN spectrometer operating at 50°C. Chemical shifts are expressed in ppm using acetone as external reference (2.223 ppm for 1H). NMR spectra were analysed using Mestrenova software.
Microtip
Manufactured by Emerson
The Microtip is a specialized laboratory equipment designed for precise and delicate sample handling. It features a compact and ergonomic design, allowing for accurate and controlled pipetting of small liquid volumes. The Microtip is intended for use in various scientific applications that require precise liquid manipulation.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using microtip
1
Molecular Mass Reduction of EPOLs
2
Transient Transfection of SERT in COS-7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
COS-7 cells were transiently transfected with SERT WT or mutants using the Lipo2000 transfection protocol (Invitrogen): 2.6 µg SERT plasmid and 7.6 µL Lipofectamine were mixed each with 200 µL Opti-MemR(1X) and incubated for 20 minutes for complex formation. The mixture was added to 10 ml DMEM 1885 medium (in house) containing 7.2 million COS-7 cells and seeded in a 150 cm2 flask. After 5 h, 20 ml DMEM1885 with Penicillin, Streptomycin, and l -glutamine was added. After 72 h, the cells were harvested for membranes by adding 10 mL PBS + 5 mM EDTA, washed with 2 mL sucrose buffer (SB: 0.3 M sucrose, 120 mM NaCl, 5 mM KCl, 1.2 mM MgSO4, 1.2 mM CaCl2, 25 mM HEPES, pH 7.4), and lysed with one ultrasonic burst (Branson Sonifier with microtip) in 1 mL SB. Membranes were pelleted at 4500xG for 20 min, resuspended in 1 mL SB, and stored at −20 °C.
3
Structural Characterization of EPOL Kp113
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The EPOL Kp113 was dissolved in water (1 g/L) and sonicated using a Branson sonifier equipped with a microtip at 2.8 Å, in order to decrease its molecular mass. The sample was cooled in an ice bath and sonicated using 5 bursts of 1 min each, separated by 1 min intervals. Polysaccharides were exchanged two times with 99.9% D 2 O by lyophilisation and then dissolved in 0.7 mL of 99.96% D 2 O.
Spectra were recorded on a 500 MHz VARIAN spectrometer operating at 50 °C for EPOL Kp113 solution and at 60°C for the polysaccharide isolated from flocs. 1D TOCSY were recorded at 50 °C using 120 ms of spin-lock time. 2D experiments were performed using standard VARIAN pulse sequences and pulsed field gradients for coherence selection when appropriate. HSQC spectra were recorded using 140 Hz (for directly attached 1 H -13 C correlations). HMBC experiments were recorded using a coupling constant of 8 Hz (for long range 1 H -13 C correlations). TOCSY spectra were acquired using 100 ms spin-lock time and 1.2 s relaxation time. NOESY experiments were recorded with 200 ms mixing time and 1.5 s relaxation time. Chemical shifts are expressed in ppm using acetone as internal reference (2.225 ppm for 1 H and 31.07 ppm for 13 C ). NMR spectra were processed using MestreNova software.
Spectra were recorded on a 500 MHz VARIAN spectrometer operating at 50 °C for EPOL Kp113 solution and at 60°C for the polysaccharide isolated from flocs. 1D TOCSY were recorded at 50 °C using 120 ms of spin-lock time. 2D experiments were performed using standard VARIAN pulse sequences and pulsed field gradients for coherence selection when appropriate. HSQC spectra were recorded using 140 Hz (for directly attached 1 H -13 C correlations). HMBC experiments were recorded using a coupling constant of 8 Hz (for long range 1 H -13 C correlations). TOCSY spectra were acquired using 100 ms spin-lock time and 1.2 s relaxation time. NOESY experiments were recorded with 200 ms mixing time and 1.5 s relaxation time. Chemical shifts are expressed in ppm using acetone as internal reference (2.225 ppm for 1 H and 31.07 ppm for 13 C ). NMR spectra were processed using MestreNova software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!