Human/Rat GFAP Antibody

(2 citations)   
  • Species Reactivity
    Human, Rat
  • Specificity
    Detects human and rat GFAP in Western blots.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human GFAP
    Leu292-Met432
    Accession # P14136
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Simple Western
    0.1-2 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Rat GFAP by Western Blot. Western blot shows lysates of rat cortical stem cells, rat brain tissue, human brain (cortex) tissue, and human brain (hypothalamus) tissue. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for GFAP at approximately 35-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
GFAP in Rat Astrocytes. GFAP was detected in immersion fixed rat astrocytes using 10 µg/mL Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
beta ‑III Tubulin in Rat Cortical Neurons and GFAP in Rat Astrocytes. beta ‑III Tubulin was detected in rat cortical neurons using 5 µg/mL Mouse Anti-neuron-specific Mouse beta ‑III Tubulin Monoclonal (clone TuJ‑1) Antibody (Catalog # MAB1195). GFAP was detected in rat astrocytes using 10 µg/mL Sheep Anti-Human GFAP Antigen Affinity-purified Poly­clonal Antibody (Catalog # AF2594). Cells were incubated with primary antibodies for 3 hours at room temperature. Cells were stained for beta‑III Tubulin using the Northern­Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and for GFAP using the Northern­Lights 493-conjugated Anti-Sheep IgG Secondary Antibody (green; Catalog # NL012). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
GFAP in Rat Cortical Stem Cells. GFAP was detected in immersion fixed 7 days differentiated rat cortical stem cells using Sheep Anti-Human GFAP Antigen Affinity-purified Poly­clonal Antibody (Catalog # AF2594) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern­Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (yellow; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human GFAP by Simple WesternTM. Simple Western lane view shows lysates of human brain (cerebellum) tissue, loaded at 0.2 mg/mL. A specific band was detected for GFAP at approximately 51 kDa (as indicated) using 0.1 µg/mL of Sheep Anti-Human/Rat GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Rat GFAP by Simple WesternTM. Simple Western lane view shows lysates of rat brain tissue, loaded at 0.2 mg/mL. A specific band was detected for GFAP at approximately 55 kDa (as indicated) using 2 µg/mL of Sheep Anti-Human/Rat GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: GFAP

GFAP (Glial fibrillary acidic protein) is a type III intermediate filament protein. It is the major component of astrocyte intermediate filament. Defects in GFAP are a cause of Alexander disease. Alexander disease is a rare disorder of the central nervous system. It is a progressive leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers which are cytoplasmic inclusions in astrocytes. At the amino acid sequence level, human GFAP shares 91% and 90% identity with rat and mouse GFAP, respectively.

  • Long Name:
    Glial Fibrillary Acidic Protein
  • Entrez Gene IDs:
    2670 (Human); 14580 (Mouse); 24387 (Rat)
  • Alternate Names:
    FLJ45472; GFAP astrocytes; GFAP; glial fibrillary acidic protein
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Dose-dependent changes in neuroinflammatory and arachidonic acid cascade markers with synaptic marker loss in rat lipopolysaccharide infusion model of neuroinflammation.
    BMC Neurosci, 2012;13(0):50.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  2. TNFalpha-induced AMPA-receptor trafficking in CNS neurons; relevance to excitotoxicity?
    Authors: Leonoudakis D, Braithwaite SP, Beattie MS, Beattie EC
    Neuron Glia Biol., 2004;1(3):263-273.
    Species: Rat
    Sample Type: Whole Cells
    Application: ICC
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