Mice were deeply anesthetized with isoflurane and perfused transcardially with 4% paraformaldehyde in 0.1 M sodium phosphate buffer. Brains were post-fixed for 1–3 days, sectioned coronally (50 μm in thickness) using a vibratome (Leica; VT1000S) and processed for immunofluorescence staining for tyrosine hydroxylase (Millipore; AB152, 1:2000), Gat1 (Cell Signaling; 37342, 1:200), Aldh1a1 (R&D Systems; AF5869, 1:200), HA (Abcam; ab18181, 1:500) or Cre (Synaptic Systems; 257 004, 1:500). After primary antibodies incubation (4°C, overnight), the following secondary antibodies (all from Thermo Fisher Scientific) were applied for 2 h at room temperature: Goat anti-mouse IgG Alexa Fluor 568 (A11004), Goat anti-mouse IgG Alexa Fluor 488 (A11029), Goat anti-rabbit IgG Alexa Fluor 488 (A11034), Goat anti-rabbit IgG Alexa Fluor 568 (A11036), Donkey anti-goat IgG Alexa Fluor 568 (A11057), Donkey anti-rabbit IgG Alexa Fluor 488 (A21206). Brain sections were mounted on Superfrost slides and coverslipped with DAPI Fluoromount-G (SouthernBiotech; 0100-20).
Af5869
Manufactured by Abcam
AF5869 is a laboratory instrument for performing Western blot analysis. Its core function is to separate and detect proteins in a sample using electrophoresis and antibody-based detection.
Automatically generated - may contain errors
2 protocols using af5869
1
Immunofluorescence Staining of Mouse Brain
2
Immunofluorescence Staining of Mouse Brain
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Mice were deeply anesthetized with isoflurane and perfused transcardially with 4% paraformaldehyde in 0.1 M sodium phosphate buffer. Brains were post-fixed for 1–3 days, sectioned coronally (50 μm in thickness) using a vibratome (Leica; VT1000S) and processed for immunofluorescence staining for tyrosine hydroxylase (Millipore; AB152, 1:2000), Gat1 (Cell Signaling; 37342, 1:200), Aldh1a1 (R&D Systems; AF5869, 1:200), HA (Abcam; ab18181, 1:500) or Cre (Synaptic Systems; 257 004, 1:500). After primary antibodies incubation (4°C, overnight), the following secondary antibodies (all from Thermo Fisher Scientific) were applied for 2 h at room temperature: Goat anti-mouse IgG Alexa Fluor 568 (A11004), Goat anti-mouse IgG Alexa Fluor 488 (A11029), Goat anti-rabbit IgG Alexa Fluor 488 (A11034), Goat anti-rabbit IgG Alexa Fluor 568 (A11036), Donkey anti-goat IgG Alexa Fluor 568 (A11057), Donkey anti-rabbit IgG Alexa Fluor 488 (A21206). Brain sections were mounted on Superfrost slides and coverslipped with DAPI Fluoromount-G (SouthernBiotech; 0100-20).
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