All oligonucleotides were synthesized, purified, and purchased from Integrated DNA Technologies (Coralville, IA). The sequences used are enumerated in Table S1 and their designs were assisted with tools in NUPACK (www.nupack.org ) and DINAMelt (http://unafold.rna.albany.edu/ ). All enzymes, proteins, and the NEBuffer 3.1 were procured from New England Biolabs (Ipswich, MA). Glycerol and dNTP mix were purchased from Promega Corporation (Madison, WI). SYBR™ Gold Nucleic Acid Gel Stain (10,000x concentrate in DMSO) was obtained from Thermo Fisher Scientific Inc. (Waltham, MA). CoOOH nanoflakes were synthesized and purified in-house. Single-donor serum was purchased from Innovative Research Inc. Gold nanospheres were purchased from nanoComposix (San Diego, CA). Graphene oxide sheets were obtained from the Nanotechnology Health Implications Research Consortium established by the National Institute of Environmental Health Sciences (NIEHS)[41 (link),42 ]. All dilutions were prepared using ultrapure water unless otherwise stated.
Gold nanospheres
Manufactured by NanoComposix
Sourced in United States
Gold nanospheres are spherical nanoparticles composed of gold. They have a typical diameter range of 5-100 nanometers. Gold nanospheres exhibit unique optical properties due to their size and shape.
Automatically generated - may contain errors
Lab products found in correlation
Sybr gold nucleic acid gel stain, by Thermo Fisher Scientific (1 mentions)
Nebuffer 3, by New England Biolabs (1 mentions)
Glycerol, by Promega (1 mentions)
Dntp mix, by Promega (1 mentions)
Hitrap protein g affinity column, by GE Healthcare (1 mentions)
Gold nanoshells, by NanoComposix (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Uranyl acetate, by Electron Microscopy Sciences (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
Ammonium hydroxide, by Avantor (1 mentions)
Thioflavin t, by Merck Group (1 mentions)
Tris base, by Merck Group (1 mentions)
Aβ1 42, by Bachem (1 mentions)
Silver nanospheres, by NanoComposix (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
N hydroxysuccinimide, by Merck Group (1 mentions)
2 n morpholino ethanesulfonic acid, by Merck Group (1 mentions)
3 n morpholino propanesulfonic acid, by Merck Group (1 mentions)
2 n morpholino ethanesulfonic acid sodium salt, by Merck Group (1 mentions)
5 protocols using gold nanospheres
1
Oligonucleotide Design and Synthesis for Nanocomposites
2
Gold-Conjugation of FMDV Monoclonal Antibody
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The pan-serotype FMDV monoclonal antibody (mAb F21-42 [24 (link)]) was purified using a Hi-Trap Protein-G affinity column (GE, Fairfield, CT, USA) and an AKIA chromatography system.
The antibody was gold-conjugated using the High Sensitivity Conjugation kit (80 nm Gold Nanospheres and 150 nm Gold Nanoshells, nanoComposix, Inc., San Diego, CA, USA). Briefly, 70 µg 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 140 µg Sulfo-NHS were added to 1 mL gold solution and incubated for 30 min at room temperature to activate carboxy gold. The gold solution was washed twice with 1 mL reaction buffer (potassium pH 7.4). The purified mAb F21-42 (20 µg) was added to the gold solution and incubated for 1 h at room temperature, followed by washing with reaction buffer twice. The gold-conjugated mAb F21-42 was re-suspended in the conjugation buffer (PBS with 0.5% BSA, 0.5% casein, 1% Tween-20 and 0.05% NaN3) and stored at 4 °C.
The antibody was gold-conjugated using the High Sensitivity Conjugation kit (80 nm Gold Nanospheres and 150 nm Gold Nanoshells, nanoComposix, Inc., San Diego, CA, USA). Briefly, 70 µg 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 140 µg Sulfo-NHS were added to 1 mL gold solution and incubated for 30 min at room temperature to activate carboxy gold. The gold solution was washed twice with 1 mL reaction buffer (potassium pH 7.4). The purified mAb F21-42 (20 µg) was added to the gold solution and incubated for 1 h at room temperature, followed by washing with reaction buffer twice. The gold-conjugated mAb F21-42 was re-suspended in the conjugation buffer (PBS with 0.5% BSA, 0.5% casein, 1% Tween-20 and 0.05% NaN3) and stored at 4 °C.
3
Amyloid-β Aggregation Inhibition
4
Nanoparticle Surface Modification Protocol
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Gold nanospheres (80 nm in diameter) and silver nanocubes (55 nm in size) were purchased from nanoComposix. Similar to gold nanocubes, the citrate-capped Gold nanospheres and polyvinyl pyrrolidone–capped silver nanocubes underwent a surfactant exchange with a mixture of 0.1 wt % of Triton X-45 and 1 mM SDS in DI water before assembly.
5
Immobilization of Antibody on Gold Nanoparticles
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N-(3-dimethylaminopropyl)-N′-carbodiimide (EDC, 97%), N-hydroxysuccinimide
(NHS, 98%), 3-(N-morpholino)propanesulfonic acid
(MOPS, 99.5%), 3-(N-morpholino)propanesulfonic acid
sodium salt (99.5%), 2-(N-morpholino)ethanesulfonic
acid (MES, 99.5%), 2-(N-morpholino)ethanesulfonic
acid sodium salt (99%), hydroxylamine 50 weight % in H2O, isopropyl alcohol (IPA, 98%) were purchased from Sigma-Aldrich
(St. Louis, MO). Hydrochloric Acid, 10% v/v aqueous solution (HCl),
was purchased from Alfa Aesar (Ward Hill, MA). Gold nanospheres (40
nm, BioReady, carboxyl, 20 OD, 5 mL, water) were obtained from NanoComposix
(San Diego, CA). A rabbit polyclonal antibody against L. monocytogenes (PIPA130487) was purchased from
Fisher Scientific and used without further purification. All reagents
were used as received. Ultrapure type-1 water (18 MΩ cm) was
acquired from an Elga PURELAB purification system and was used for
all buffer preparations. Thermo Scientific Pierce 96-Well Plates,
Product No. 15041 was used for bicinchoninic acid (BCA) assay. Bovine
serum albumin (BSA) standard kit was purchased from Thermo Fisher
catalog #23225.
(NHS, 98%), 3-(N-morpholino)propanesulfonic acid
(MOPS, 99.5%), 3-(N-morpholino)propanesulfonic acid
sodium salt (99.5%), 2-(N-morpholino)ethanesulfonic
acid (MES, 99.5%), 2-(N-morpholino)ethanesulfonic
acid sodium salt (99%), hydroxylamine 50 weight % in H2O, isopropyl alcohol (IPA, 98%) were purchased from Sigma-Aldrich
(St. Louis, MO). Hydrochloric Acid, 10% v/v aqueous solution (HCl),
was purchased from Alfa Aesar (Ward Hill, MA). Gold nanospheres (40
nm, BioReady, carboxyl, 20 OD, 5 mL, water) were obtained from NanoComposix
(San Diego, CA). A rabbit polyclonal antibody against L. monocytogenes (PIPA130487) was purchased from
Fisher Scientific and used without further purification. All reagents
were used as received. Ultrapure type-1 water (18 MΩ cm) was
acquired from an Elga PURELAB purification system and was used for
all buffer preparations. Thermo Scientific Pierce 96-Well Plates,
Product No. 15041 was used for bicinchoninic acid (BCA) assay. Bovine
serum albumin (BSA) standard kit was purchased from Thermo Fisher
catalog #23225.
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