One milliliter of resuspended pSGNs was injected into a 1-cm light-path cuvette. The absorption spectrum of pSGNs was scanned using a UV-visible spectrophotometer. The morphology of the pSGNs was observed using a TEM. Ten microliters of pSGNs were loaded onto carbon-coated grids and placed on top of Watman paper to absorb excess solvent. The TEM images were recorded with a JEOL JEM-1200EXII operating at the bias voltage of 80 kV. To determine the particle concentration, the absorption of pSGNs was first adjusted to OD800 = 6 and subsequently serially diluted in distilled water. Next, the diluted pSGNs were mixed with 1.85 × 106/mL of 3-μm latex beads (Sigma, St. Louis, MO) at a ratio of 1:1 and evenly separated on 0.1-μm filter membranes (Millipore, Tullagreen, Ireland) through suction. Next, the membranes were coated with platinum and observed using an SEM (s-3500N, Hitachi, Tokyo, Japan). The SEM images were analyzed using ImageJ software to count the ratio of the pSGNs and 3-μm latex beads.
3 μm latex beads
Manufactured by Merck Group
Sourced in Australia
3-μm latex beads are spherical particles made of a synthetic polymer material. They have a diameter of approximately 3 micrometers and are commonly used in various laboratory applications, such as calibration, flow cytometry, and as a reference standard.
Automatically generated - may contain errors
2 protocols using 3 μm latex beads
1
Characterization of Plasmonic Gold Nanoparticles
2
Antibody Validation and Cell Labeling
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Rab31 antibody was purchased from Abnova and APPL2 antibody from Abcam. Phospho-Akt (Ser-473), phosphoxp38 MAPK (Thr-180/Tyr-182), and Rab5 antibodies used in this study were purchased from Cell Signaling Technology (Beverly, MA). GST antibody was purchased from Invitrogen (Mulgrave, VIC, Australia). Anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was purchased from Trevigen (Gaithersburg, MD). LAMP1 antibody was purchased from BD Biosciences (San Jose, CA). Alexa Fluor 488–, 594–, and 647–conjugated secondary antibodies were purchased from Molecular Probes/Invitrogen (Eugene, OR). Horseradish peroxidase–conjugated goat anti-mouse and anti-rabbit antibodies were obtained from Zymed (San Francisco, CA). Bacterial lipopolysaccharide (LPS), purified from Salmonella enterica serotype Minnesota Re 595, was purchased from Sigma-Aldrich (Castle Hill, NSW, Australia). For phagocytosis, human IgG (Invitrogen) was conjugated to 3-μm latex beads (Sigma-Aldrich) and sheep red blood cells (Australian Ethical Biologicals, Coburg, Australia) was conjugated with rabbit anti-sheep IgG (Sigma-Aldrich). All other chemicals and reagents were from Sigma-Aldrich.
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