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Antibodies against gfap

Manufactured by Cell Signaling Technology
Sourced in United States

Antibodies against GFAP are laboratory reagents used to detect and quantify the presence of the Glial Fibrillary Acidic Protein (GFAP) in biological samples. GFAP is an intermediate filament protein found in astrocytes and other glial cells in the central nervous system. These antibodies can be used in various immunodetection techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and distribution of GFAP in different cell types and tissue samples.

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2 protocols using antibodies against gfap

1

Biochemical Markers Quantification Protocol

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FITC-dextran (MilliporeSigma, Burlington, MA, USA), ELISA kits for TNF-α, IL-1β, IL-10, and lipocalin were purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against GFAP, Aβ, BACE-1, and PSD-95 were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Anti-synaptophysin, anti-synapsin 1, anti-cFos, and anti-COX-2 antibodies were purchased from Abcam (Cambridge, MA, USA). Anti-Iba-1 antibody was purchased from Wako Chemicals USA, Inc. (Richmond, VA, USA). iNOS antibody was purchased from BD, Biosciences (San Jose, CA, USA). Anti-GAPDH, α-tubulin antibodies and horseradish peroxidase-conjugated secondary antibodies (goat anti-mouse IgM, goat anti-mouse IgG, goat anti-rabbit, goat anti-rat, bovine anti-goat and bovine anti-mouse were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Elite Vectastain ABC reagents, Vector VIP, biotinylated anti-rabbit, and anti-mouse antibodies were purchased from Vector Laboratories Inc (Burlingame, CA, USA). Standards used in mass spectroscopy analysis were purchased from MilliporeSigma (Burlington, MA, USA). Autosampler vials were obtained from ThermoFisher Scientific, (Waltham, MA, USA), silanized micro-vial inserts were from Agilent (Santa Clara CA; part #5181-8872) and inserts were from VWR (Radnor, PA, USA).
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2

Quantifying Astrocytic Activation in Mouse Brain

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Mice were anesthetized with pentobarbital (80 mg/kg, intraperitoneal injection) and perfused via the aorta with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO) in PBS. Brains were removed and postfixed for 24 h in the same fixative at 4°C. Brains were immersed in 30% sucrose for 24 h, frozen, and cut in 35 μm sections using a cryostat (Leica, Germany). Free-floating sections were incubated in 50% alcohol for 20 min, followed by 10% normal donkey serum in PBS for 30 min, and then antibodies against GFAP (1:100; Cell Signaling, Danvers, MA) overnight. Sections were then incubated in 2% normal donkey serum in PBS for 10 min followed by Alexa 488-conjugated secondary goat anti-mouse antibody (1:1000; Cell Signaling, Danvers, MA) for 2 h. Images from each brain region of interest (CPu, NAc core, NAc shell) were obtained using an LSM 510 confocal laser scanning microscope (Carl Zeiss, Germany). Areas of GFAP and GFAP-EGFP-positive astrocytes within regions of interest (450 μm × 450 μm) were quantified using NIH Image J software (Bethesda, MD).
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