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.
Amicon ultra 4 ym 100 centrifuge membrane concentrators
Manufactured by Merck Group
The Amicon Ultra 4 YM-100 centrifuge membrane concentrators are a laboratory equipment designed for sample concentration. The device uses a semipermeable membrane to separate and concentrate specific molecules or macromolecules from a solution through centrifugation.
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2 protocols using amicon ultra 4 ym 100 centrifuge membrane concentrators
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.
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