Five µm frozen section of tumor samples were permeabilized with ethanol-acetone (19v–1v) before saturation in PBS- BSA5%. Sequential staining with anti-CD31/anti-MMP9 antibodies or anti-CD45/anti-MMP9 antibodies were done (CD31, JC70A, Dako, 1/50—CD45, HI30, Invitrogen, 1/200—MMP9, ab38898 Abcam, 1/1000). Secondary antibodies were purchased from Molecular Probes™, goat anti mouse-AlexaFluor 488 (A11001) for antibodies targeting CD31 and CD45 and goat anti rabbit-AlexaFluor 568 (A11011) for anti-MMP9 antibody. Nuclei were stained with Hoechst 33,342 (B2261, Sigma, 1/1000). Fluorescent microcopy pictures were done with Zeiss® Observer. Z1 and ZenPRO software and analysis were done with ImageJ software.
Goat anti rabbit alexafluor 568 a11011
Manufactured by Thermo Fisher Scientific
Goat anti-rabbit AlexaFluor 568 (A11011) is a secondary antibody conjugated with the AlexaFluor 568 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in immunoassays and other applications.
Automatically generated - may contain errors
2 protocols using goat anti rabbit alexafluor 568 a11011
1
Multicolor Immunofluorescence Imaging of Tumor
2
Immunohistochemistry of Adult Brain Samples
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Adult brains were dissected in PBS and fixed in 4% paraformaldehyde for 20 min at room temperature. Samples were washed three times in PBT (phosphate buffered saline with 1% Triton X-100 at pH 7.2) and blocked with 5% normal goat serum for one hour. Samples were then incubated with primary antibodies overnight at 4°C. After three additional washes with PBT, samples were incubated with a secondary antibody overnight at 4°C. Stained brains were mounted in Vectashield (Vector Laboratories, Burlingame, CA, USA) and recorded with confocal fluorescence laser scanning microscopy (Zeiss). Anti-Crz antibody (1:1000) and goat anti-rabbit-Alexa Fluor 568 (A-11011, Molecular Probes, 1:200 dilution) were used. GtACR1::YFP and GtACR2::YFP were visualized without antibody staining. Control mCD8::GFP expression was visualised using anti-GFP (ab13970) and anti-chicken-Alexa Fluor 488 antibody staining.
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