Paraffin-embedded brain tissues were sliced into 6 μm sections, and then subjected to immunofluorescence staining. After 5 min of antigen retrieval, the sections were incubated in 5% goat serum for 1 h and incubated with rabbit TH (dilution ratio of 1:500, GB11181, Servicebio, Wuhan, China) and 4-HNE (dilution ratio of 1:50, MA5-27570, Invitrogen), and α-SYN (dilution ratio of 1:500, ab272736, Abcam) at 4 °C overnight and with Cy3-conjugated donkey anti-rabbit antibody (dilution ratio of 1:250, GB 21403, Servicebio) or Goat Anti-Mouse Alexa Fluor 488 (dilution ratio of 1:1000, A32723, Invitrogen) for 1 h. A confocal laser microscope (Olympus) was employed to observe the immune-response cells.
Gb21403
Manufactured by Wuhan Servicebio Technology
Sourced in China
GB21403 is a laboratory equipment product. It is designed for specialized scientific applications. The core function of this equipment is to perform specific tasks within a controlled laboratory environment. No further details on the intended use or application of this product can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Gb11181, by Wuhan Servicebio Technology (2 mentions)
Ab272736, by Abcam (1 mentions)
A confocal laser microscope, by Olympus (1 mentions)
Ma5 27570, by Thermo Fisher Scientific (1 mentions)
A32723, by Thermo Fisher Scientific (1 mentions)
Laser confocal microscope, by Olympus (1 mentions)
Cm1950, by Leica camera (1 mentions)
Ab185628, by Abcam (1 mentions)
Ab7800, by Abcam (1 mentions)
Dapi gdp1024, by Wuhan Servicebio Technology (1 mentions)
Cy3 conjugated donkey anti rabbit igg, by Wuhan Servicebio Technology (1 mentions)
Gb25301, by Wuhan Servicebio Technology (1 mentions)
Nile red, by Merck Group (1 mentions)
Laser scanning confocal microscope, by Nikon (1 mentions)
Tissue tek optimum cutting temperature compound, by Sakura Finetek (1 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (1 mentions)
Ab97931, by Abcam (1 mentions)
Gb22404, by Wuhan Servicebio Technology (1 mentions)
Gb13063, by Wuhan Servicebio Technology (1 mentions)
5 protocols using gb21403
1
Immunofluorescence Staining of Paraffin-Embedded Brain Tissues
2
Tracking Exosome Trafficking in Parkinson's
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To verify whether Exos can reach the SN through the BBB, PD rats were randomly divided into two groups (n = 3 per group): 6-OHDA+saline and 6-OHDA+Exos groups. The procedure for labeling Exos with PKH67 is described above. After 24 h, animals were anaesthetized and their brains were harvested and post-fixed for 1 day with 4% paraformaldehyde. They were then transferred to 10%, 20%, and 30% sucrose solutions at 4 °C, then embedded in optimal cutting temperature (OCT) compound. Brains were sectioned at 6 μm thickness using a microtome (CM1950; Leica, Heerbrugg, Switzerland). After washing with PBS, sections were incubated with 1% bovine serum albumin containing 0.3% Triton X-100 in PBS. Sections were then incubated with rabbit TH (dilution 1:500, GB11181, Servicebio, Wuhan, China) at 4 °C overnight. The next day, sections were incubated with Cy3-conjugated donkey anti–rabbit antibody (dilution 1:250, GB21403, Servicebio) at room temperature for 1 h. Each of the steps was followed by three 5-min rinses in PBS. Immunoreactive cells were visualized using the confocal laser microscope (Olympus).
3
Immunofluorescence Analysis of Mouse Meibomian Glands
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The isolated MG tissues were snap-frozen in Tissue-Tek optimum cutting temperature compound (Sakura Finetek, Tokyo, Japan), cut into 8-µm-thick sections, and mounted on poly-L-lysine–coated glass slides. The MGs sections or cultured cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.5% Triton X-100 for 30 minutes, and blocked with 5% BSA for at least 1 hour at room temperature and then incubated with rabbit monoclonal antibody to keratin 6 (Krt6) (1:200, ab93279; Abcam, Cambridge, UK), Krt1 (1:200, ab185628; Abcam, Cambridge, UK) and mouse monoclonal antibody to Krt6 (1:200, ab218438; Abcam, Cambridge, UK), PPARγ (1:200, sc-7273, Arigo, Hamburg, Germany), and keratin 14 (Krt14) (1:200, ab7800; Abcam, Cambridge, UK) overnight at 4°C. After incubation, the samples were stained Cy3-conjugated Donkey Anti-Rabbit IgG (1:500, GB21403; Servicebio, Wuhan, China) or Alexa Fluor 488, Goat Anti-Mouse IgG (1:500, GB25301; Servicebio, Wuhan, China) at 37°C for 1 hour. DAPI (GDP1024; Servicebio, Wuhan, China), Nile Red (72485; Sigma-Aldrich, St. Louis, MO, USA), and LipdTOX (H34477; Thermo Fisher, Waltham, MA, USA) were used for nucleus and lipid staining, respectively. Images were obtained with a laser-scanning confocal microscope (Nikon, Tokyo, Japan).
4
Immunofluorescence Staining of RASAL2 Protein
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Immunofluorescence staining was performed on cells cultured on cover slips. The cells were washed twice with PBS, fixed with 4% paraformaldehyde and permeabilized with 0.25% Triton X-100. Slides were blocked with 5% BSA and stained with anti-RASAL2 primary antibody (1:300; 22140; Proteintech, Wuhan, China) followed by incubation with Cy3 conjugated secondary antibody (1:200; GB21403; Servicebio, Wuhan, China) and BODIPY493/503 (1 µg/mL; Thermo Fisher Scientific). 4′,6-diamidino-2-phenylindole was used to counterstain the nuclei. The slides were visualized and photographed with a fluorescence microscope (Nikon, Tokyo, Japan).
5
Immunofluorescence Analysis of Epiretinal Membranes
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Epiretinal membranes were cut in 4-µm sections and fixed on slides. To remove paraffin, the slides were immersed in xylene twice for 15 minutes and then in graded ethanol (5 minutes in 100% ethanol, 5 minutes in 85% ethanol, and 5 minutes in 75% ethanol). After being washed with distilled water, the slides were immersed in Tris-EDTA buffer (pH = 8.0) in a microwave for antigen retrieval. The slides were washed three times with PBS after cooling. A 3% BSA solution was used to block the sections for 30 minutes at room temperature. The diluted primary antibody was incubated with sections in a wet box at 4°C overnight. The corresponding secondary antibodies and 4′,6-diamidino-2-phenylindole (DAPI) for nucleus staining were incubated the next day. Images were captured under confocal microscopy (Leica, Wetzlar, Germany). The antibodies involved were as follows: anti-UCP2 (ab97931, 1:400; Abcam), anti-SIRT3 (#2627, 1:400; Cell Signaling Technology), anti-CD31 (GB13063, 1:100; Servicebio, Wuhan, China), anti-goat FITC (GB22404, 1:100; Servicebio), and anti-rabbit Cyanine3 (GB21403, 1:100; Servicebio).
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