Sections were blocked with 10% normal goat serum (NGS) in PBS with 0.1% Triton X-100 (PBS-T) for 1 h. The following primary antibodies were incubated at 4°C overnight in PBS-T and 5% NGS: anti-glial fibrillary acidic protein (GFAP), either mouse monoclonal (1:2000, Millipore) or chicken polyclonal (1:400, abcam) for astrocytes; anti-vimentin, mouse monoclonal (1:200, Sigma) for reactive astrocytes; anti-NeuN, mouse monoclonal (1:200, Chemicon) for mature neurons; anti-Iba-1 rabbit polyclonal (1:750, Wako) for microglia; and anti-collagen IV rabbit polyclonal (1:3000, Chemicon) a component of the basal lamina that is used as a specific marker for cerebral microvessels (15 (link)). Sections were washed in PBS-T three times and incubated with the corresponding Alexa Fluor 568-conjugated (red) and Alexa Fluor 488-conjugated (green) IgG secondary antibodies (all 1:1000, Invitrogen) for 2 h at room temperature. Sections were rinsed with PBS and distilled water and coverslipped with ProLong Gold antifade reagent with DAPI (Invitrogen).
Alexa fluor 568 conjugated red
Manufactured by Thermo Fisher Scientific
Alexa Fluor 568-conjugated (red) is a fluorescent dye used in various biological applications. It has an excitation maximum of 578 nm and an emission maximum of 603 nm, producing a red fluorescent signal. The dye is commonly used for labeling and detection purposes in techniques such as immunofluorescence, flow cytometry, and microscopy.
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Lab products found in correlation
Anti neun, by Merck Group (2 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)
Anti iba 1 rabbit polyclonal, by Fujifilm (1 mentions)
Anti vimentin, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Anti iba1, by Fujifilm (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Anti cd31, by BD (1 mentions)
2 protocols using alexa fluor 568 conjugated red
1
Immunohistochemical Labeling of Neural Cells
2
Immunohistochemical Staining of Neural Markers
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Sections were washed three times in phosphate-buffered saline with 0.1% TritonÔ X-100 (PBST) for 5 min each, and blocked with 10% normal goat serum (NGS) in PBST for 1 h. The following primary antibodies were incubated at 4°C overnight in PBST/5% NGS: anti-glial fibrillary acidic protein (GFAP), chicken polyclonal (1:1000; Abcam; Ab4647); anti-Iba1, mouse monoclonal (1:1000; Wako; 019-19741); anti-NeuN, mouse monoclonal (1:1000; Millipore; NG1876252); or anti-CD31, rat monoclonal (1:300; BD Biosciences; 550274). Sections were washed three times in PBST and incubated with either Alexa Fluor 568-conjugated (red), Alexa Fluor Ò 488-conjugated (green), or Alexa Fluor 647-conjugated (far-red) IgG secondary antibody (1:1000, Invitrogen) overnight at 4°C. Sections were then rinsed with PBST and deionized water, mounted on gelatin coated slides, covered with ProLong Gold antifade reagent with 4¢,6-diamidino-2-phenylindole (Invitrogen), and protected with cover-slips.
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