Anti-GFAP antibody - Astrocyte Marker (ab7260)
Overview
- Product nameAnti-GFAP antibody - Astrocyte MarkerSee all GFAP primary antibodies ...
- DescriptionRabbit 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.
Some customers have successfully used ab7260 on Zebrafish lysates; however we have conflicting data to suggest that not all batches will be suitable for work on Zebrafish. For further information, please contact Abcam Scientific Support.
- Tested applicationsIHC-FoFr, IHC-Fr, ICC/IF, WB, IHC-P, ICC more details
- Species reactivityReacts 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 notesIn some cases, the antibody may appear red in color. This is due to small amounts of hemolysis, and does not affect antibody performance.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.01% Sodium Azide
- PurityWhole antiserum
- Clonality Polyclonal
- IsotypeIgG
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Research Areas
Applications
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.
| Application | Notes |
|---|---|
| IHC-FoFr | IHC-FoFr: 1/5000. See Abreview. |
| IHC-Fr | IHC-Fr: 1/500. |
| ICC/IF | ICC/IF: 1/1000. |
| WB | WB: 1/50000. Detects a band of approximately 55,48 kDa. This lower 48kDa band is thought to be a degradation product. |
| IHC-P | IHC-P: 1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| ICC | ICC: 1/5000. |
Target
- FunctionGFAP, 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 specificityExpressed in cells lacking fibronectin.
- Involvement in diseaseDefects 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 similaritiesBelongs to the intermediate filament family.
- Post-translational
modificationsPhosphorylated by PKN1. - Cellular localizationCytoplasm. Associated with intermediate filaments.
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Database links
- Entrez Gene: 281189 Cow
- Entrez Gene: 2670 Human
- Entrez Gene: 14580 Mouse
- Entrez Gene: 24387 Rat
- Omim: 137780 Human
- SwissProt: Q28115 Cow
- SwissProt: P14136 Human
- SwissProt: P03995 Mouse
- SwissProt: P47819 Rat
- Unigene: 514227 Human
- Unigene: 1239 Mouse
- Unigene: 91512 Rat
see all
Target information above from: UniProt accession
P14136
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- Astrocyte antibodyFLJ42474 antibodyFLJ45472 antibody
- GFAP antibodyGFAP_HUMAN antibodyGlial Fibrillary Acidic Protein antibodyIntermediate filament protein antibody
see all
Anti-GFAP antibody - Astrocyte Marker images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)Carl Hobbs, King`s College London, United KingdomImmunohistochemistical 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.
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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.
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Immunohistochemistry (Frozen sections) - GFAP antibody - Astrocyte Marker (ab7260)This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemyab7260 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. -
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)This image is courtesy of an Abreview submitted by Mr Osama Mohsenab7260 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.
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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 minuteThis image is courtesy of an anonymous Abreview
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)This image was kindly supplied by Dr Vladimir Milenkovic by Abreviewab7260 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. -
Immunocytochemistry/ Immunofluorescence - GFAP antibody - Astrocyte Marker (ab7260)This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemyab7260 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.
References for Anti-GFAP antibody - Astrocyte Marker (ab7260)
This product has been referenced in:
- Valverde AM et al. Proapoptotic and survival signaling in the neuroretina at early stages of diabetic retinopathy. Mol Vis 19:47-53 (2013). WB ; Human . Read more (PubMed: 23335850) »
- Albert-Weissenberger C et al. Blocking of bradykinin receptor B1 protects from focal closed head injury in mice by reducing axonal damage and astroglia activation. J Cereb Blood Flow Metab : (2012). IHC-Fr ; Mouse . Read more (PubMed: 22569191) »
