In brief, RP membrane (Millipore) was striped using a noncontact BioJet HR value with a high-resolution syringe pump attached to an XYZ3050 platform (BioDot, CA) with the 3D6 capture mAb as a test line (T) and a donkey-anti-mouse IgG used for the control line (C). The RP membranes were water washed, then blocked in polyvinylpyrrolidone (PVP40; Sigma) and dried. The 4C9 mAb was conjugated to 40 nm gold (InnovaCoat Gold; Innova Biosciences) and 10 OD sprayed onto a 10 mm glass fiber conjugate pad (Millipore) using a noncontact AirJet HR aerosol dispenser (BioDot) attached to the XYZ platform. Dried membranes were adhered to 60 mm plastic backing card with 25 mm Fusion-5 membrane (GE Healthcare) as a sample pad and 22 mm CF6 membrane (Millipore) as an absorbent sink. The test strips were cut (60 × 4.5 mm) and housed in a two-part plastic cassette with a pressure point at material overlap. Dilutions of SEB were added to the sample pad (100 μL) and resolved for 10 minutes and then photographed.
Innovacoat gold
Manufactured by Abcam
InnovaCoat GOLD is a lab equipment product designed for the functionalization of gold nanoparticles. It provides a simple and effective way to attach a wide range of ligands, including proteins, peptides, oligonucleotides, and small molecules, to the surface of gold nanoparticles.
Automatically generated - may contain errors
3 protocols using innovacoat gold
1
Rapid Lateral Flow Immunoassay for SEB
2
Labeling and Inoculation of Viruses
3
Labeling and Inoculation of Viruses
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mice were infected via footpad using an insulin syringe and injected the specified doses of VACV or MVA from purified stocks (in a total volume of 10 μls) into wild-type C57Bl/6 animals. VACVs used in this study were VV-NP-S-GFP, MVA-NP-S-GFP, and VACV-NP-eGFP (an identical virus lacking the SIINFEKL determinant). All viruses have been previously described and virus stocks were grown and titered in house, as described 10 (link), 52 (link). ZIKV was grown and tittered in Vero cells. 104 FFU of 104 pfu of ZIKV H/PF/2013 was inoculated footpad in 10 μls into Ifnar−/− mice.
Alexa-647 labeled virus was prepared with sucrose-purified virus and a Protein Labeling Kit (ThermoFisher) according to the manufacturer’s instructions. Labeled virus was purified twice through a spin column to remove free dye and then by sucrose gradient. Gold-labeled VACV was prepared using purified virus and a was conjugated to 10 nm gold nanoparticle conjugation kit (InnovaCoat GOLD, Innova Biosciences). Virus and reaction buffer were mixed according to the manufacturer’s instructions and the mixture was used to reconstitute InnovaCoat GOLD particles. The reaction mixture was incubated at room temperature for 15 min, quenched for 5 min at 25 °C, spun down and resuspended in 1mM TRIS pH 9.0 buffer prior to injection.
Alexa-647 labeled virus was prepared with sucrose-purified virus and a Protein Labeling Kit (ThermoFisher) according to the manufacturer’s instructions. Labeled virus was purified twice through a spin column to remove free dye and then by sucrose gradient. Gold-labeled VACV was prepared using purified virus and a was conjugated to 10 nm gold nanoparticle conjugation kit (InnovaCoat GOLD, Innova Biosciences). Virus and reaction buffer were mixed according to the manufacturer’s instructions and the mixture was used to reconstitute InnovaCoat GOLD particles. The reaction mixture was incubated at room temperature for 15 min, quenched for 5 min at 25 °C, spun down and resuspended in 1mM TRIS pH 9.0 buffer prior to injection.
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