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Anti gfap primary antibody

Manufactured by Abcam
Sourced in United States

Anti-GFAP primary antibody is a laboratory reagent that specifically binds to the Glial Fibrillary Acidic Protein (GFAP), a key intermediate filament protein found in astrocytes and other glial cells. This antibody can be used in various immunological techniques to detect and analyze the presence and distribution of GFAP in biological samples.

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2 protocols using anti gfap primary antibody

1

Western Blot Analysis of GFAP Expression

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Fresh coronal slices containing caudal NTS (300-µm thick) were prepared from 300–500 g male rats (4 rats/group) and incubated with 100 µM FC or Veh for 2 h (the same procedure as for mIPSCs recording). Caudal NTS was dissected from surrounding tissue and each was separately homogenized in the ice-cold Radio-Immunoprecipitation Assay homogenization buffer, sonicated, and centrifuged at 30,000 g for 20 min. Supernatant was collected and protein concentration was measured by bicinchoninic acid assay (Pierce, Cat. No. 23225). Protein (25 µg) from each sample was mixed with 6 µL of SDS-loading buffer dye and boiled for 5 min. Protein samples were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membranes. After blocking with 5% nonfat milk at room temperature, membranes were incubated with rabbit polyclonal anti-GFAP primary antibody (1:5,000, Abcam, ab7260, Waltham, MA) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit IgG secondary antibody (1:10,000, Invitrogen, No. 31460) for 1 h at room temperature. Blots were visualized by enhanced chemiluminescence. Norm-Veh and CIH-Veh were set to 100%. Changes of GFAP/β-actin GeneTex, (GTX629630, Irvine, CA) ratio as a function of FC (100 µM) incubation were compared with their vehicle control and shown as a percentage of control intensity.
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2

Immunocytochemical Analysis of Neural Cells

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Differentiated and undifferentiated cells were fixed by incubation in 4% paraformaldehyde/PBS, were permeated with Triton X-100 (0.4%) in PBS for 5 min and processed for ICC using 5% goat serum (Sigma, USA) (23 (link)), anti-NSE primary antibody (1:100, Santa Cruz, USA), anti-β-III-tubulin primary antibody (1:50, Santa Cruz, USA), and anti-GFAP primary antibody (1:200, Abcam, USA) to confirm the expression of neural-specific proteins by ICC. The affinity purified goat Anti-Mouse IgG Rhodamine (TRITC) conjugated antibody was used as the secondary antibody (1:200, Millipore, USA), and the nuclei were stained with 4’, 6-diamidino-2-phenylindole (DAPI; 1:500, Roche) and visualized under a microscope.
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