Cell cultures were washed once in PBS and fixed in 4% formaldehyde solution at 4°C for 45 min. To remove the fixative, cells were washed in PBS four times. Next, PBS containing 5% albumin and 0.05% saponin was used to reduce unspecific binding and to permeabilize the cells before immunostaining with a mouse monoclonal anti-HIV-1-p24 antibody (0.5 μg/ml) (Abcam) for 2 h at room temperature, followed by three washes and a secondary staining for 1 h using goat anti-mouse conjugated with Alexa Fluor 568 (Abcam). Finally, the cells were washed and mounted using ProLong Gold Antifade Mountant with DAPI (Life Technologies).
Goat anti mouse conjugated with alexa fluor 568
Manufactured by Abcam
Sourced in United Kingdom
Goat anti-mouse antibody conjugated with Alexa Fluor 568 dye. This product is used for detection and visualization in various immunoassays and fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Mouse monoclonal anti hiv 1 p24 antibody, by Abcam (1 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Mouse anti iba1, by Abcam (1 mentions)
Blocking buffer, by Beyotime (1 mentions)
Rabbit anti inos, by Abcam (1 mentions)
Mouse anti brdu, by Abcam (1 mentions)
Rabbit anti neun, by Abcam (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Lmw cs, by Merck Group (1 mentions)
Dylight 488 nhs ester, by Thermo Fisher Scientific (1 mentions)
Label it sirna tracker kit, by Mirus Bio (1 mentions)
3 protocols using goat anti mouse conjugated with alexa fluor 568
1
Immunostaining of HIV-1-p24 in Fixed Cells
2
Immunofluorescent Staining of Brain Slices
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Using prepared brain tissue slices for immunofluorescent staining. The brain slice was obstructed using a blocking buffer (Beyotime Biotechnology, China), supplemented with primary antibodies, and left to incubate overnight at 4 ℃. Subsequently, secondary antibodies were introduced and incubated for 6 h at room temperature in a darkroom. The main antibodies employed were mouse anti-Iba-1 (at a dilution of 1:800, bought from Abcam in the United Kingdom), rabbit anti-INOS (at a dilution of 1:400, also from Abcam in the United Kingdom), rabbit anti-NeuN (at a dilution of 1:400, from Abcam), and mouse anti-BrdU (at a dilution of 1:200, bought from Abcam).The secondary antibodies used were goat anti-rabbit conjugated with Alexa Fluor® 488 (at a dilution of 1:1000, obtained from Abcam) and goat anti-mouse conjugated with Alexa Fluor® 568 (at a dilution of 1:800, obtained from Abcam).Additionally, Hoechst 33342 (at 1:2000, from ThermoFisher Scientific, Inc.) was employed to counterstain cell nuclei for half an hour at room temperature in a light-protected environment. Using a fluorescence microscope (Leica DMR, from Germany), positive cells were observed.
3
Chitosan Molecular Weight Influences
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Two
types of chitosan were used in this study.
The low molecular weight chitosan (LMW CS) had a viscosity average
molecular weight of 50 kDa, and the degree of deacetylation was 75–85%;
a higher molecular weight chitosan (HMW CS) had a viscosity average
molecular weight of 60–120 kDa, and the degree of deacetylation
was 80%. LMW CS and acetic acid glacial were purchased from Sigma-Aldrich
Inc. (St. Louis, MO), and HMW CS was a kindly provided by KitoZyme
S.A. (Belgium). Fluorescent dye DyLight-488 NHS ester and succinimidyl-([N-maleimidopropionamido]ethylene glycol)ester (NHS-PEG-MAL)
were purchased from Thermo Scientific (Rockford, IL). Double-stranded
siRNAs against Mad2 and nontargeting (NT) siRNA sequence were purchased
from Santa Cruz Biotechnology Inc. Amine-reactive Alexa Fluor 488
was purchased from Invitrogen/Life Technologies (Carlsbad, CA), and
a Label IT siRNA tracker kit was procured from Mirus Corporation (Madison,
WI). Pico-Green fluorescence reagent for quantification of double-stranded
nucleic acid constructs was purchased from Invitrogen/Life Technologies
(Carlsbad, CA). For Mad2 protein quantification by flow cytometry,
the antibody monoclonal anti-Mad2 clone 17D10 was obtained from Sigma
(Sigma-Aldrich, St. Louis, MO), and the secondary antibody goat anti-mouse
conjugated with Alexa Fluor 568 was from Abcam (Abcam, Cambridge,
UK).
types of chitosan were used in this study.
The low molecular weight chitosan (LMW CS) had a viscosity average
molecular weight of 50 kDa, and the degree of deacetylation was 75–85%;
a higher molecular weight chitosan (HMW CS) had a viscosity average
molecular weight of 60–120 kDa, and the degree of deacetylation
was 80%. LMW CS and acetic acid glacial were purchased from Sigma-Aldrich
Inc. (St. Louis, MO), and HMW CS was a kindly provided by KitoZyme
S.A. (Belgium). Fluorescent dye DyLight-488 NHS ester and succinimidyl-([N-maleimidopropionamido]ethylene glycol)ester (NHS-PEG-MAL)
were purchased from Thermo Scientific (Rockford, IL). Double-stranded
siRNAs against Mad2 and nontargeting (NT) siRNA sequence were purchased
from Santa Cruz Biotechnology Inc. Amine-reactive Alexa Fluor 488
was purchased from Invitrogen/Life Technologies (Carlsbad, CA), and
a Label IT siRNA tracker kit was procured from Mirus Corporation (Madison,
WI). Pico-Green fluorescence reagent for quantification of double-stranded
nucleic acid constructs was purchased from Invitrogen/Life Technologies
(Carlsbad, CA). For Mad2 protein quantification by flow cytometry,
the antibody monoclonal anti-Mad2 clone 17D10 was obtained from Sigma
(Sigma-Aldrich, St. Louis, MO), and the secondary antibody goat anti-mouse
conjugated with Alexa Fluor 568 was from Abcam (Abcam, Cambridge,
UK).
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