Serial, frozen sections (40 μm, coronal) were prepared as described above. For immunofluorescence staining of DCX and GFAP, free floating sections were incubated for 3 h in a blocking buffer consisting of 10% normal donkey serum and 0.3% Triton X-100 in PBS and subsequently overnight in 1% BSA, 0.3% Triton X-100 in PBS containing either a goat anti-DCX (Santa Cruz #SC8066; 1:250) or a rabbit anti-GFAP (Sigma #180063; 1:1000), respectively. After rinsing with PBS, the sections were blocked in the aforementioned blocking buffer for 3 h and incubated in the dark for 6 h with either secondary Alexa Fluor-594 donkey anti-goat IgG antibody (Life Technologies; 1:1000) or secondary Alexa Fluor-594 donkey anti-rabbit IgG antibody (Life Technologies; 1:1000). The sections were then rinsed in PBS. After the final PBS rinse, the sections were mounted on SuperfrostPlus slides (Fisher), allowed to dry at RT in the dark, coverslipped and stored at -20°C. The sections were imaged with Olympus IX81/DSU spinning disc confocal microscope.
Alexa fluor 594 donkey anti goat igg antibody
Manufactured by Thermo Fisher Scientific
Sourced in Canada
Alexa Fluor-594 donkey anti-goat IgG antibody is a fluorescently labeled secondary antibody used for the detection and visualization of goat primary antibodies in various immunodetection techniques, such as immunohistochemistry, immunocytochemistry, and Western blotting.
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Lab products found in correlation
Ix 81 dsu spinning disc confocal microscope, by Olympus (1 mentions)
Alexa fluor 594 donkey anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
Superfrost plus slide, by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
Sc 507, by Santa Cruz Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Santa Cruz Biotechnology (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
2 protocols using alexa fluor 594 donkey anti goat igg antibody
1
Immunofluorescence of Neuronal and Glial Markers
2
Immunofluorescence Analysis of ASC Cytokine Expression
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To analyze the ASC cytokine expression, ASCs were cultured on the coverslips (VWR International) until confluence. The ASCs were then fixed with 4% paraformaldehyde (Alfa Aesar, Ward Hill, MA, USA) in PBS for 1 h, and treated with 0.1% Triton X-100 (Sigma-Aldrich) for 15 min. After blocking with 3% bovine serum albumin (BSA, Sigma-Aldrich) in PBS at room temperature for 1 h, the slides were incubated with rabbit polyclonal anti-vascular endothelial growth factor (VEGF) antibody (1:100 dilution, sc-507; Santa Cruz Biotechnology), goat polyclonal anti-insulin-like growth factor-1 (IGF-1) antibody (1:100 dilution, sc-1422; Santa Cruz Biotechnology), or rabbit polyclonal anti-hepatocyte growth factor (HGF) antibody (1:100 dilution, sc-7949; Santa Cruz Biotechnology) at 4°C overnight. After incubation with secondary antibody Alexa Fluor 594 goat anti-rabbit IgG antibody (1:200 dilution, A11012; Life Technologies) or Alexa Fluor 594 donkey anti-goat IgG antibody (1:200 dilution, A11058; Life Technologies) at room temperature for 1 h, cell nuclei were counterstained with 4¢,6-diamidino-2-phenylindole dihydrochloride (DAPI; Santa Cruz Biotechnology).
The images were taken with a fluorescence microscope (Zesis Axio observer Z1 Inverted microscope; Carl Zesis Canada Ltd., Toronto, ON, Canada).
The images were taken with a fluorescence microscope (Zesis Axio observer Z1 Inverted microscope; Carl Zesis Canada Ltd., Toronto, ON, Canada).
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