HMEC-1 cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton-X-100. The proliferation was visualized using bromodeoxyuridine (BrdU) staining. Slides were stained with anti-BrdU (1:100, ABclonal, Wuhan, China, #A20304), followed by Cy3-conjugated anti-rabbit antibody (1:200, Invitrogen, Carlsbad, CA, USA, #A27039). CD31 staining was performed on 5 μm thick Matrigel plug slides. Slides were stained with anti-CD31 (1:100, Santa Cruz Biotechnology, Santa Cruz, CA, USA, #Sc-376764), followed by Cy3-conjugated anti-mouse antibody (1:200, Invitrogen, #A21424). For functional vessel identification, mice were injected with FITC-labeled isolectin-B4 via tail vein, 30 min before sacrifice. Double IF staining was conducted on 5 μm thick gastrocnemius muscles. The perfused capillaries were labeled with anti-CD31 (1:100, Santa, #Sc-376764), followed by Cy3-conjugated anti-mouse antibody (1:200, Invitrogen, #A21424) and FITC-conjugated anti-rabbit antibody (1:200, Abcam, MA, USA, #AB6717). The density of small arteries (CD31+/α-SMA+) and capillaries (CD31+/α-SMA−) were double stained with anti-CD31 (1:100, Santa, #Sc-376764) and anti-α-SMA (1:100, ABclonal, A17910), followed by Cy3-conjugated anti-mouse antibody (1:200, Invitrogen, #A21424) and FITC-conjugated anti-rabbit antibody (1:200, Abcam, #AB6717).
Fitc conjugated anti rabbit antibody
Manufactured by Abcam
Sourced in United States
FITC-conjugated anti-rabbit antibodies are secondary antibodies that are conjugated with the fluorescent dye fluorescein isothiocyanate (FITC). These antibodies are used to detect and visualize primary antibodies that have been raised in rabbit, in a variety of immunochemical applications such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Vimentin, by Abcam (2 mentions)
Confocal microscope, by Olympus (2 mentions)
Cy3 conjugated anti mouse antibodies, by Abcam (2 mentions)
E cadherin, by Abcam (2 mentions)
Anti mouse cy3 conjugated antibody, by Thermo Fisher Scientific (1 mentions)
Anti cd31, by Santa Cruz Biotechnology (1 mentions)
Prolong antifade reagent, by Thermo Fisher Scientific (1 mentions)
Lsm 510 meta confocal microscope, by Zeiss (1 mentions)
Spss 13, by IBM (1 mentions)
Dako fluorescence mounting medium, by Agilent Technologies (1 mentions)
5 protocols using fitc conjugated anti rabbit antibody
1
Visualization of Cellular Proliferation and Vascular Remodeling
2
Immunofluorescence Analysis of Cellular Markers
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An immunofluorescence analysis was performed on 8-μm-thick frozen sections, which were fixed with ice-cold 4% paraformaldehyde for 15 minutes and blocked with normal serum for 20 minutes at room temperature before being incubated with one or more specific antibodies against vimentin (1:200, Abcam, Cambridge), E-cadherin (1:200, Abcam, Cambridge), Sufu (1:200, Abcam, Cambridge) or Gli-1 (1:200, Abcam, Cambridge) overnight and in the dark at 4°C. After three washes, the slides were then stained with FITC-conjugated anti-rabbit antibodies or Cy3-conjugated anti-mouse antibodies (1:500, Abcam, Cambridge). The nuclei were counterstained with DAPI. Stained cells were visualized with an Olympus confocal microscope. All of the experiments were repeated at least three times.
3
Immunofluorescence Localization of Viral Capsids
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HeLa cells attached in a monolayer to cover slips were fixed with 4% paraformaldehyde (PFA) for 10 h at 4 °C. Then cover slips with cells were incubated with 0.1% Triton X-100 solution in 1× PBS for 7 min to improve antibody penetration. To block nonspecific antibody binding, cells were further incubated with blocking solution containing 4% BSA in 1× PBS for 24 hr at 4 °C. The previously described rabbit antibodies recognizing the epitope in the common C-terminal of BgDV1 capsid proteins or rabbit antibodies against GFP (Abcam, Cambridge, MA, USA), both at 1:1000 dilution, were used as the primary antibodies. The incubation with primary antibodies was performed for 2 hr with subsequent washing with 1× PBS. The FITC-conjugated anti-rabbit antibodies (Abcam, USA) were used as the secondary antibodies. Secondary antibodies were diluted 1:5000, and incubation was performed also for 2 hr with subsequent washing with PBS. The slides were mounted with ProLong Antifade Reagent (Invitrogen, USA) containing DAPI, and the cell nuclei were allowed to stain for several hours in the dark. The intracellular localization of the fluorescent signals was analyzed in a Carl Zeiss LSM 510 Meta confocal microscope.
4
Immunofluorescence and Western Blot Analysis
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Immunofluorescence. The immunofluorescence analysis was performed on 8-µm-thick frozen sections of tissue, which were fixed with ice-cold 4% paraformaldehyde for 15 min and blocked with normal serum for 20 min at room temperature before being incubated with one or more specific antibodies against vimentin (1:200), E-cadherin (1:200), SUFU (1:200) or TWIST1 (1:200) (all from Abcam, Cambridge, UK) overnight and in the dark at 4˚C. After three washes, the slides were stained with FITC-conjugated anti-rabbit antibodies or Cy3-conjugated anti-mouse antibodies (1:500; Abcam). The nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI). Stained cells were visualized with an Olympus confocal microscope (Olympus Corp., Tokyo, Japan). All the experiments were repeated at least three times.
Western blot analysis. All the cell lysates were prepared and western blot analysis was performed as previously described (20) .
The following antibodies were used: E-cadherin ( Table II. Association between TWIST1 expression in LAD patients' characteristics. -------------------------------------------------------------------------- one-way ANOVA test. A difference was considered to be statistically significant at P<0.05. All the statistical analyses were performed with the SPSS 13.0 software (IBM Corp., Armonk, NY, USA).
Western blot analysis. All the cell lysates were prepared and western blot analysis was performed as previously described (20) .
The following antibodies were used: E-cadherin ( Table II. Association between TWIST1 expression in LAD patients' characteristics. -------------------------------------------------------------------------- one-way ANOVA test. A difference was considered to be statistically significant at P<0.05. All the statistical analyses were performed with the SPSS 13.0 software (IBM Corp., Armonk, NY, USA).
5
Immunohistochemistry of Adipose Tissue
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Adipose tissues were fixed in 4% paraformaldehyde, prepared as paraffin blocks, and sectioned at 8 µm. Paraffin blocks were deparaffinized and rehydrated in alcohol. Antibody against target protein was incubated overnight. After washing the primary antibody, FITC‐conjugated anti‐rabbit antibody (Abcam, USA) was incubated for 2 h. Sealing‐stained slides were prepared by mounting a coverslip with a Dako Fluorescence Mounting Medium (DAKO, Denmark) and scanned with AxioScan.Z1 (Zeiss, Germany).
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