The largest database of trusted experimental protocols

10nm gold particles

Manufactured by Aurion
Sourced in United States

10nm gold particles are a type of laboratory equipment used for various scientific applications. These particles have a diameter of 10 nanometers and are composed of the metallic element gold. They are commonly used in fields such as nanotechnology, biotechnology, and materials science for their unique optical and physical properties.

Automatically generated - may contain errors

10 protocols using 10nm gold particles

Electron microscopy was performed as described [13 (link)]. Briefly, exosome pellets obtained from serum were suspended in 100 μl of PBS, loaded onto Formwar carbon-coated grids and contrasted with 2% uranyl acetate. For immunogold staining the grids were placed into a blocking buffer step for 1 hr. Without rinsing, the grids were immediately placed into the primary antibody at the appropriate dilution overnight at 4°C (monoclonal anti-human-CD36 1:10, clone 99.1, bioceros; monoclonal anti-CD63 1:10, Abcam). As controls, some grids were not exposed to the primary antibody. The next day the grids were rinsed with PBS then floated on drops of IgA protein attached with 10nm gold particles (AURION, Hatfield, PA) for 1 hour at room temperature. Grids were rinsed with PBS and were placed in 1% Glutaraldehyde in 0.1M Phosphate buffer for 15 minutes. After rinsing in PBS and distilled water the grids were contrasted and embedded in a mixture of 2% methyl cellulose and 1% uranyl acetate. The grids were examined with a Tecnai G2 Spirit transmission electron microscope (TEM) (FEI Europe, Eindhoven, The Netherlands) and images were recorded using a Morada CCD camera (Olympus Soft Image Solutions GmbH, Münster, Germany).
+ Open protocol
+ Expand
Vero E6 cells were fixed for at least 1 h with glutaraldehyde 2.5% in 0.1M sodium cacodylate buffer. Cells were then incubated with anti-vimentin (V9) antibody (Santa Cruz Biotechnology) followed by a secondary goat anti-mouse IgG (H + L) antibody coupled to 10 nm-gold particles (Aurion). For resin embedding, cells were washed three times with a mixture of 0.2M saccharose/0.1M sodium cacodylate. Cells were post-fixed for 1 h with 1% OsO4 diluted in 0.2M Potassium hexa-cyanoferrate (III)/0.1M sodium cacodylate solution. After three 10 min washes with distilled water, the cells were gradually dehydrated with ethanol by successive 10 min baths in 30, 50, 70, 96, and 100% ethanol. Substitution was achieved by successively placing the cells in 25, 50, and 75% Epon solutions for 15 min. Cells were placed for 1 h in 100% Epon solution and in fresh Epon 100% over-night under vacuum at room temperature. Polymerization occurred with cells in 100% fresh Epon for 72 h at 60°C. Ultrathin 70 nm sections were cut using a UC7 ultramicrotome (Leica) and placed on HR25 300 Mesh Copper/Rhodium grids (TAAB, United Kingdom). Sections were contrasted according to Reynolds.60 (link) Images were obtained by Tecnai G2 TEM (Thermo-Fischer/FEI) operated at 200 keV equipped with a 4096 × 4096 pixels’ resolution Eagle camera (FEI).
+ Open protocol
+ Expand
Fixed specimens at an optimal concentration were placed onto a 400 mesh carbon/formvar coated grids and allowed to absorb to the formvar for a minimum of 1 min. The grids were placed into a blocking buffer for a block/permeabilization step for one hour. Without rinsing, the grids were immediately placed into the primary antibody at the appropriate dilution overnight at 4 °C (Mouse anti-PD-L1, sc-293425, Santa Cruz). As controls, some of the grids were not exposed to the primary antibody. The following day, all the grids were rinsed with PBS then floated on drops of the appropriate secondary antibody attached with 10 nm gold particles (AURION, Hatfield, PA) for two hours at room temperature. Grids were rinsed with PBS and were placed in 2.5% Glutaraldehyde in 0.1 M Phosphate buffer for 15 min. After rinsing in PBS and distilled water, the grids were allowed to dry and stained for contrast using uranyl acetate. The samples were viewed with a Tecnai Bio Twin transmission electron microscope (FEI, Hillsboro, OR) and images were taken with an AMT CCD Camera (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
Protein extracts from mouse brains, PNC, OC and HEK293 cells were applied on 300-mesh carbon-coated grids in drops of 3 µL for 5 min, blotted on a filter paper and air-dried before the immunogold procedure. Grids were blocked in PBS, 0.1 % cold water fish gelatin (PBS-CWFG) for 5 min before incubating with PBS-CWFG diluted primary antibodies (HT7 1:50, AT8 1:50, AT100 1:50) for 90 min at RT. After washing 5 times (2 min) in PBS-CWFG, grids were incubated for 60 min with secondary antibody (goat anti-mouse IgG) labeled with 10-nm gold particles (Aurion) diluted 1:30 in PBS-CWFG. Following washes in PBS and dH2O, grids were negatively stained with 2 % uranyl acetate for 1 min. The grids were examined with a JEOL JEM1400 transmission electron microscopy equipped with a Olympus SIS Quemesa 11 Mpxl camera, and images were taken at magnifications of 20× and 30× k (resp. pixel size = 0.72 and 0.48 nm). Immuno-gold electron-microscopy on brain slices is described in detail in supplemental data.
+ Open protocol
+ Expand
Fixed specimens at an optimal concentration were placed onto a 300 mesh
carbon/formvar coated grids and allowed to absorb to the formvar for a minimum
of 1 minute. For immunogold staining the grids were placed into a blocking
buffer for a block/permeabilization step for 1 hour. Without rinsing, the grids
were immediately placed into the primary antibody at the appropriate dilution
overnight at 4°C (monoclonal anti-CD9 1:10, Abcam). As controls, some
grids were not exposed to the primary antibody. The next day, all of the grids
were rinsed with PBS then floated on drops of the appropriate secondary antibody
attached with 10nm gold particles (AURION, Hatfield, PA) for 2 hours at room
temperature. Grids were rinsed with PBS and were placed in 2.5%
Glutaraldehyde in 0.1M phosphate buffer for 15 minutes. After rinsing in PBS and
distilled water the grids were allowed to dry and stained for contrast using
uranyl acetate. The samples were viewed with a Tecnai Bio Twin transmission
electron microscope (FEI, Hillsboro, OR) and images were taken with an AMT CCD
Camera (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
The grids were placed into blocking buffer for 1 h and then into a solution of the primary antibody at the appropriate dilution overnight at 4 °C (anti-CD151, 96283S, CST; 1:500 dilution). Subsequently, PBS was used to rinse the grids, which were floated on drops of the secondary antibody attached to 10-nm gold particles (AURION) at room temperature for 1.5 h. The grids were rinsed with PBS and placed into 2.5% glutaraldehyde in 0.1 M phosphate buffer for 15 min. Finally, the grids were stained with uranyl acetate and viewed under a Tecnai G2 20 Twin transmission electron microscope (FEI; 200 kV).
+ Open protocol
+ Expand
Fixed specimens were placed on a 400-mesh carbon/Formvar coated grid at optimal concentration and allowed to absorb into the formvar for 1 min. For immunogold staining, the raster was placed in blocking buffer for 1 h in the blocking/permeation step. Without rinsing, the raster was immediately added to the primary antibody dilution (1:300 anti-CD9 ab92726, Abcam and anti-GPC1 PIPA528055, Thermo Fisher Scientific) overnight at 4 °C. As a control, some grids were not exposed to the primary antibody. The next day, the grids were washed with PBS and then floated in 10 nm gold particles (Aurion, Hatfield, PA, USA) ligated with appropriate secondary antibodies and incubated at room temperature for 2 h. They were then rinsed with PBS and placed in 2.5% glutaraldehyde in 0.1 M phosphate buffer for 15 min. The grids were rinsed in PBS and distilled water, dried, and stained with uranyl acetate. Samples were photographed with a Tecnai Bio Twin transmission electron microscope (FEI, Hillsboro, OR, USA) and images were taken with an AMT CCD camera (Advanced Microscopy Technology, Danvers, MA, USA).
+ Open protocol
+ Expand
Transmission
electron microscopy (TEM) characterization was performed using a JEOL
JEM-2100F field emission S/TEM equipped with an Oxford X-MaxN 80 mm2
SDD detector for elemental analysis. TEM images were acquired at a
200 kV accelerating voltage and 116 μA emission current. DigitalMicrograph
GMS3 (Gatan) and Aztec TEM (Oxford Instruments) software packages
were used for the TEM data and energy dispersive X-ray (EDX) spectral
analysis and interpretation, respectively. Briefly, for the immunogold
labeling with antibodies, GPC-1 antibodies (pa5–51290, Thermo
Scientific) were attached on 10 nm gold particles (AURION) according
to the manufacturer’s instructions at room temperature. Healthy
plasma and PDAC plasma sample pools were prepared. 200 μL from
each of 5 healthy or 5 PDAC plasma samples was taken to form a healthy
plasma or PDAC plasma sample pool. Exosomes were isolated as previously
described. Then isolated exosome samples were conjugated with the
gold-particles attached GPC-1 antibodies at the appropriate dilution
overnight at 4 °C. Samples for TEM characterization were drop-cast
from dilute aqueous suspensions onto amorphous carbon coated (200
nm) Cu grids (Agar Scientific) and dried naturally overnight.
+ Open protocol
+ Expand
Fixed specimens at an optimal concentration were placed onto a 300 mesh
carbon/formvar coated grids and allowed to absorb to the formvar for a minimum
of 1 minute. For immunogold staining the grids were placed into a blocking
buffer for a block/permeabilization step for 1 hour. Without rinsing, the grids
were immediately placed into the primary antibody at the appropriate dilution
overnight at 4°C (monoclonal anti-CD9 1:10, Abcam). As controls, some
grids were not exposed to the primary antibody. The next day, all of the grids
were rinsed with PBS then floated on drops of the appropriate secondary antibody
attached with 10nm gold particles (AURION, Hatfield, PA) for 2 hours at room
temperature. Grids were rinsed with PBS and were placed in 2.5%
Glutaraldehyde in 0.1M phosphate buffer for 15 minutes. After rinsing in PBS and
distilled water the grids were allowed to dry and stained for contrast using
uranyl acetate. The samples were viewed with a Tecnai Bio Twin transmission
electron microscope (FEI, Hillsboro, OR) and images were taken with an AMT CCD
Camera (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
Fixed specimens at an optimal concentration were placed onto a 400 mesh carbon/formvar coated grids and allowed to absorb to the formvar for a minimum of 1 minute. The grids were placed into a blocking buffer for a block/permeabilization step for 1 hour. Without rinsing, the grids were immediately placed into the primary antibody at the appropriate dilution overnight at 4 °C (Mouse anti-PD-L1, sc-293425, Santa Cruz, CA, USA). As controls, some of the grids were not exposed to the primary antibody. The following day, all the grids were rinsed with PBS then oated on drops of the appropriate secondary antibody attached with 10 nm gold particles (AURION, Hat eld, PA) for two hours at room temperature. Grids were rinsed with PBS and were placed in 2.5% Glutaraldehyde in 0.1M Phosphate buffer for 15 min.
After rinsing in PBS and distilled water the grids were allowed to dry and stained for contrast using uranyl acetate. The samples were viewed with a Tecnai Bio Twin transmission electron microscope (FEI, Hillsboro, OR) and images were taken with an AMT CCD Camera (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!