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Products:Neuroscience >> Cell Adhesion Proteins >> Cytoskeletal Proteins >> Intermediate Filaments
Overview
Product name
Anti-GFAP antibody - Astrocyte Marker
See all GFAP products (35) ...
Description
Rabbit polyclonal to GFAP - Astrocyte Marker
Specificity
Specifically recognizes mammalian GFAP on western blots and immunocytochemically. Detects a band of 55kDa corresponding to GFAP and also a GFAP derived 25kDa band.
Tested applications
IHC-FoFr, IHC-Fr, ICC/IF, WB, IHC-P, ICCmore details
Cross reactivity
Reacts with
Mouse, Rat, Cat, Human, Marmoset (common)
Predicted to work with
Cow, Pig, all Mammals
Immunogen
The initial immunization was performed with a preparation of full length human recombinant GFAP expressed in bacteria and highly purified. Subsequent boosts were performed with GFAP purified from a Triton X-100 extract of myelin associated material from bovine spinal cord, following an "axonal flotation" procedure (Liem et al.). The GFAP was further purified by centrifugation and ion exchange chromatography in 6m urea on DEAE cellulose.
General notes
In some cases, the antibody may appear red in color. This is due to small amounts of hemolysis, and does not affect antibody performance.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.01% Sodium Azide
Purity
Whole antiserum
Clonality
Polyclonal
Isotype
IgG
Research areas
Stem Cells >> Neural Stem Cells >> Glial Restricted Lineage >> Astrocyte
Tags & Cell Markers >> Cell Type Markers >> Neuroscience Markers >> Glial
Signal Transduction >> Cytoskeleton / ECM >> Cytoskeleton >> Intermediate Filaments >> Class II >> GFAP
Neuroscience >> Cell Type Marker >> Glia marker >> Astrocyte marker
Neuroscience >> Cell Adhesion Proteins >> Cytoskeletal Proteins >> Intermediate Filaments
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)
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Immunocytochemistry/ Immunofluorescence - GFAP antibody - Astrocyte Marker (ab7260)
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Immunohistochemistry (Frozen sections) - GFAP antibody - Astrocyte Marker (ab7260)
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Applications
Show applications key
Our Abpromise guarantee covers the use of ab7260 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-FoFr: 1/5000(See Abreview.)
IHC-Fr: 1/500
ICC/IF: 1/1000
WB: 1/50,000Detects a band of approximately 55,48 kDa.(This lower 48kDa band is thought to be a degradation product.)
IHC-P: 1/5000Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC
ICC: 1/5000ICC: 1/5000
Target
Function
GFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells.
Tissue specificity
Expressed in cells lacking fibronectin.
Involvement in disease
Defects in GFAP are a cause of Alexander disease (ALEXD) [MIM:203450]. 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. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death usually within the first decade. Infants with Alexander disease develop a leukoencephalopathy with macrocephaly, seizures, and psychomotor retardation. Patients with juvenile or adult forms typically experience ataxia, bulbar signs and spasticity, and a more slowly progressive course.
Sequence similarities
Belongs to the intermediate filament family.
Post-translational
modifications
Phosphorylated by PKN1.
Cellular localization
Cytoplasm. Associated with intermediate filaments.
Target information above from: UniProt accessionP14136
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Astrocyte antibody
- FLJ42474 antibody
- FLJ45472 antibody
see all
Database links
- Entrez Gene: 281189 Cow
- Entrez Gene: 2670 Human
- Entrez Gene: 14580 Mouse
- Entrez Gene: 396562 Pig
- Entrez Gene: 24387 Rat
- Omim: 137780 Human
- SwissProt: Q28115 Cow
- SwissProt: P14136 Human
see all
Anti-GFAP antibody - Astrocyte Marker images:
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)
Immunohistochemistical detection of GFAP antibody - Astrocyte Marker (ab7260) on formaldehyde-fixed paraffin-embedded monkey brain sections. Antigen retrieval step: Heat mediated. Buffer Used: Citric acid pH6. Permeabilization: None. Blocking step: 1% BSA for 10 mins @ 21°C. Primary antibody incubated at 1/2000 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti rabbit IgG Conjugated to biotin (1/200). Marmoset brain: astrocytes are clearly and strongly labelled.
Carl Hobbs, King`s College London, United Kingdom
Immunocytochemistry/ Immunofluorescence - GFAP antibody - Astrocyte Marker (ab7260)
ab7260 at 1/10000 dilution staining Mouse cortical astrocytes by Immunocytochemistry. The cells were permeabilized with Triton/HEPES buffer prior to primary application. The antibody was incubated with the cells for 18 hours and then bound antibody was detected with an Alexa Fluor ® 488 conjugated goat anti-rabbit antibody.
This image is courtesy of an Abreview submited by Charmaine Noonan.
Immunohistochemistry (Frozen sections) - GFAP antibody - Astrocyte Marker (ab7260)
ab7260 at a 1/5000 dilution staining rat spinal cord tissue sections by IHC-Fr. Rats were transcardially perfused with 4% PFA. The tissue was post fixed 1 hour in 4% PFA and then 30% sucrose for three days. 20µm sections were cryostat cut. The primary antibody was incubated with the tissue sections for 18 hours. Bound antibody was detected using an Alexa Fluor ® 488 conjugated goat anti-rabbit polyclonal.
This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy
Western blot - GFAP antibody - Astrocyte Marker (ab7260)
Western blot of whole rat cerebellum homogenate stained with ab7260 at dilution of 1:100,000. A prominent band running with an apparent SDS-PAGE molecular weight of ~50kDa corresponds to rodent GFAP. A lower band at ~45kDa is derived from the GFAP molecule.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)
ab7260 staining rat brain tissue sections by IHC-P. Sections were fixed in formaldehyde and bocoked with a commercialy available blocking agent prior to incubating with ab7260, diluted 1/5000 for 20 hours at 4°C. A HRP conjugated mouse polyclonal (universal HRP polymer detection) antibody was used as the secondary.
This image is courtesy of an Abreview submitted by Mr Osama Mohsen
Western blot - GFAP antibody - Astrocyte Marker (ab7260)
All lanes : Anti-GFAP antibody - Astrocyte Marker (ab7260) at 1/5000 dilution
Lane 1 : Rat thoracotomy, spinal cord homogenate
Lane 2 : Rat thoracotomy, spinal cord homogenate
Lane 3 : Rat thoracotomy, spinal cord homogenate
Lane 4 : Rat thoracotomy sham, spinal cord homogenate
Lane 5 : Rat thoracotomy sham, spinal cord homogenate
Lane 6 : Rat nerve transect sham, spinal cord homogenate
Lane 7 : Rat nerve transect sham, spinal cord homogenate
Lane 8 : Rat nerve transect, spinal cord homogenate
Lane 9 : Rat nerve transect, spinal cord homogenate
Lysates/proteins at 30 µg per lane.
Secondary
HRP conjugated goat anti-rabbit at 1/3000 dilution
developed using the ECL technique
Observed band size : 53 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
This image is courtesy of an anonymous Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)
ab7260 staining GFAP in mouse eye tissue sections by Immunohistochemistry (paraffin embedded sections). Tissue was fixed with paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then permeabilized using 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C; followed by incubation with the primary antibody, at a 1/500 dilution, for 16 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 used at a 1/5000 dilution.The retinal layers are: ganglion cells layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), and photoreceptor outer segments (ROS). Nuclei were counterstained with DAPI.
This image was kindly supplied by Dr Vladimir Milenkovic by Abreview
Immunocytochemistry/ Immunofluorescence - GFAP antibody - Astrocyte Marker (ab7260)
ab7260 staining rat pup cortical preps by ICC/IF. The preps were grown for 14 days in culture and plated onto coverslips. The preps were acid/alcohol fixed and blocked prior to incubation with ab7260. Bound antibody was detected using an Alexa Fluor ®488 conjugated goat polyclonal antibody. Nuclei were visualised using DAPI.
This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy
References for Anti-GFAP antibody - Astrocyte Marker (ab7260)
This product has been referenced in:
- Kinsella Pet al. Imatinib and docetaxel in combination can effectively inhibit glioma invasion in an in vitro 3D invasion assay. J Neurooncol 101:189-98 (2011). WB; Human.Read more (PubMed: 20512610) »
- Zhao Set al. Fluorescent labeling of newborn dentate granule cells in GAD67-GFP transgenic mice: a genetic tool for the study of adult neurogenesis. PLoS One 5: (2010). IHC-P; Mouse.Read more (PubMed: 20824075) »
See all 13 publications for this product
Publishing research using ab7260? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"