Immediately after the last imaging session, mice were sacrificed and perfused with formalin, and their brains were extracted. The tissue was immediately immersed in formalin. Coronal sections (30 μm) were generated using a vibrating microtome (Fisher). For nestin, GFAP, and TUJ-1 staining, sections were incubated for 1 hour in a blocking solution (0.3% bovine serum albumin, 8% goat serum, and 0.3% Triton X-100) at room temperature, followed by incubation at 4°C overnight with the following primary antibodies diluted in blocking solution: (i) anti-human nestin (Millipore), (ii) anti-GFAP (Millipore), (iii) anti-TRAIL (Prosci), and (iv) anti–TUJ-1 (Sigma). Sections were washed three times with PBS, incubated in the appropriate secondary antibody, and visualized using a confocal microscope (Olympus).
Anti human nestin
Manufactured by Merck Group
Anti-human nestin is a laboratory reagent used for the identification and characterization of nestin, an intermediate filament protein expressed in stem and progenitor cells. It is a specific antibody that binds to nestin, enabling its detection and quantification in various experimental and analytical applications.
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Lab products found in correlation
Anti tuj1, by Merck Group (1 mentions)
Vibrating microtome, by Thermo Fisher Scientific (1 mentions)
Anti gfap, by Merck Group (1 mentions)
Alexa fluor conjugated phalloidin, by Thermo Fisher Scientific (1 mentions)
Anti human β catenin, by Abcam (1 mentions)
Col 94, by Merck Group (1 mentions)
Anti human vimentin, by Thermo Fisher Scientific (1 mentions)
Glial fibrillary acidic protein (gfap), by Abcam (1 mentions)
Matrigel, by BD (1 mentions)
2 protocols using anti human nestin
1
Immunohistochemical Analysis of Neural Markers
2
Immunocytochemical Analysis of Cell Markers
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The following antibodies were used: anti-mouse β-catenin (mouse clone 14/beta-catenin, 1:100, BD Biosciences); anti-human β-catenin (rabbit polyclonal, 1:1000, Abcam); anti-mouse N-cadherin (mouse, clone GC-4, 1:200, Sigma); anti-mouse laminin (rabbit polyclonal, 1:100, Sigma); anti-human collagen-IV (mouse, clone Col-94, 1:300, Sigma); anti-human vimentin (rabbit, SP20 clone, human specific, 1:300, Thermo Scientific), anti-bovine glial fibrillary acidic protein (GFAP) (chicken polyclonal, 1:1000, Abcam); anti-human nestin (rabbit polyclonal, human specific, 1:100, Millipore). Primary antibodies were visualized with secondary AlexaFluor-conjugated goat-anti-mouse, goat-anti-rabbit or goat-anti-chicken polyclonal antibodies (Invitrogen; 5 µg/ml). For background controls, isotype-matched unspecific mouse IgG (BD Biosciences) was used for monoclonal antibody stainings, and for polyclonal antibody stainings samples were incubated only with secondary AlexaFluor-conjugated antibodies. Cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; 2.5 µg/ml). F-actin was labelled with AlexaFluor-conjugated phalloidin (Invitrogen). Growth factor-reduced reconstituted basement membrane (rBM) (Matrigel, BD Biosciences; 9.8 mg/ml) was used for rBM interface culture.
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