The separation from free MPEG-gPLL was accomplished by loading 1 ml of AuNP3s concentrated in Amicon Ultra 4 YM-100 centrifuge membrane concentrators (EMD-Millipore, Billerica MA) on top of a step-gradient consisting of 0.2 ml 50% Opti-Prep (60% Iodixanol solution in saline, Sigma-Aldrich, St. Louis MO) with the initial density 1.32 g/ml, followed by 4 ml of 8% solution of Opti-Prep in 0.03M Hepes, pH 7.5 and centrifuged in a SW55.1 Ti rotor (Beckman) at 40,000×rpm (RCF 152,000×g) for 40 min. The bottom 0.4 ml fraction was collected, vortexed and washed with 20 mM Hepes, 0.1 M NaCl, pH 8.0 using Amicon Ultra 4 YM-100 centrifuge membrane concentrators as suggested by the manufacturer. After the washings, the particles were pooled and lyophilized. The lyophilized particles were reconstituted in water at 10 mg/ml for further experiments. Before thermogravimetric analysis, extensively dialyzed and lyophilized AuNP3 were dried under high vacuum for 3 days. The purity of nanoparticles was determined using a Superdex 200 size-exclusion HPLC column (GE-Healthcare Life Sciences) eluted with 0.1M ammonium acetate buffer, pH 7.0.
Sw55.1 ti rotor
Manufactured by Beckman Coulter
The SW55.1 Ti rotor is a high-speed ultracentrifuge rotor designed for Beckman Coulter centrifuge systems. It is constructed using titanium alloy materials, providing strength and durability. The rotor is capable of reaching high rotational speeds, enabling efficient separation of small sample volumes.
Automatically generated - may contain errors
3 protocols using sw55.1 ti rotor
1
Purification of Gold Nanoparticles
2
Purification of Gold Nanoparticles
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The separation
from free MPEG-gPLL
was accomplished by loading 1 mL of AuNP3s concentrated in Amicon
Ultra 4 YM-100 centrifuge membrane concentrators (EMD-Millipore, Billerica
MA) on top of a step-gradient consisting of 0.2 mL 50% Opti-Prep (60%
Iodixanol solution in saline, Sigma-Aldrich, St. Louis MO) with the
initial density 1.32 g/mL, followed by 4 mL of 8% solution of Opti-Prep
in 0.03 M Hepes, pH 7.5, and centrifuged in a SW55.1 Ti rotor (Beckman)
at 40 000 rpm (RCF 152 000 g) for 40
min. The bottom 0.4 mL fraction was collected, vortexed, and washed
with 20 mM Hepes, 0.1 M NaCl, pH 8.0 using Amicon Ultra 4 YM-100 centrifuge
membrane concentrators as suggested by the manufacturer. After the
washings, the particles were pooled and lyophilized. The lyophilized
particles were reconstituted in water at 10 mg/mL for further experiments.
Before thermogravimetric analysis, extensively dialyzed and lyophilized
AuNP3 were dried under high vacuum for 3 days. The purity of nanoparticles
was determined using a Superdex 200 size-exclusion HPLC column (GE-Healthcare
Life Sciences) eluted with 0.1 M ammonium acetate buffer, pH 7.0.
from free MPEG-gPLL
was accomplished by loading 1 mL of AuNP3s concentrated in Amicon
Ultra 4 YM-100 centrifuge membrane concentrators (EMD-Millipore, Billerica
MA) on top of a step-gradient consisting of 0.2 mL 50% Opti-Prep (60%
Iodixanol solution in saline, Sigma-Aldrich, St. Louis MO) with the
initial density 1.32 g/mL, followed by 4 mL of 8% solution of Opti-Prep
in 0.03 M Hepes, pH 7.5, and centrifuged in a SW55.1 Ti rotor (Beckman)
at 40 000 rpm (RCF 152 000 g) for 40
min. The bottom 0.4 mL fraction was collected, vortexed, and washed
with 20 mM Hepes, 0.1 M NaCl, pH 8.0 using Amicon Ultra 4 YM-100 centrifuge
membrane concentrators as suggested by the manufacturer. After the
washings, the particles were pooled and lyophilized. The lyophilized
particles were reconstituted in water at 10 mg/mL for further experiments.
Before thermogravimetric analysis, extensively dialyzed and lyophilized
AuNP3 were dried under high vacuum for 3 days. The purity of nanoparticles
was determined using a Superdex 200 size-exclusion HPLC column (GE-Healthcare
Life Sciences) eluted with 0.1 M ammonium acetate buffer, pH 7.0.
3
Ultracentrifugal Separation of MPEG5-gPLL Au-IO
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