Recombinant Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (ab279290)
![Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] (ab279290)](https://www.abcam.com/ps/products/279/ab279290/Images/ab279290-458721-anti-gfap-antibody-immunocytochemistry-neural-glia-mouse.jpg "Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] (ab279290)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [EPR1034Y] to GFAP - Mouse IgG2a
- Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra), ICC
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a
See all GFAP primary antibodies -
Description
Mouse monoclonal [EPR1034Y] to GFAP - Mouse IgG2a -
Host species
Mouse -
Tested applications
Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra), ICCmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human cerebral cortex tissue. WB: Human, mouse and rat brain tissue lysate. IHC-Fr: Mouse cerebrum tissue. Flow Cyt (intra): Rat primary neural/glia cells. IP: Rat brain tissue lysate. ICC: mouse neural/glia cells
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General notes
This mouse antibody has been engineered from a RabMAb parent antibody (ab68428). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1034Y -
Isotype
IgG2a -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab279290 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000.
|
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| IHC-Fr |
1/500.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
|
| IP |
1/30.
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| IHC-P |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Flow Cyt (Intra) |
1/1000.
|
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| ICC |
1/50.
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| Notes |
|---|
|
WB
1/1000. |
|
IHC-Fr
1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
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IP
1/30. |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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Flow Cyt (Intra)
1/1000. |
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ICC
1/50. |
Target
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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
modificationsPhosphorylated by PKN1. -
Cellular localization
Cytoplasm. Associated with intermediate filaments. - Information by UniProt
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Database links
- Entrez Gene: 2670 Human
- Entrez Gene: 14580 Mouse
- Entrez Gene: 24387 Rat
- Omim: 137780 Human
- SwissProt: P14136 Human
- SwissProt: P03995 Mouse
- SwissProt: P47819 Rat
- Unigene: 514227 Human
see all -
Alternative names
- wu:fb34h11 antibody
- ALXDRD antibody
- cb345 antibody
see all
Images
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Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] (ab279290)
Immunocytochemical analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized mouse neural/glia tissue labeling GFAP with ab279290 at 1/50 (16 µg/ml) dilution followed by ab98736 Goat Anti-Mouse IgG (DyLight® 488) pre-adsorbed at 1/1000 dilution (Green). Positive staining on mouse primary astrocytes is observed. The nuclear counterstain was DAPI (Blue).
Counterstain: ab207165 anti-GFAP rabbit mAb at 1/1000 dilution with secondary antibody ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution.
Negative controls: ab279290 with secondary ab150080 at 1/500 dilution; and ab207165 at 1/1000 dilution with secondary ab98376 at 1/1000 dilution
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![Western blot - Anti-GFAP antibody [EPR1034Y] (ab279290)](https://www.abcam.com/ps/products/279/ab279290/Images/ab279290-450219-anti-gfap-antibody-epr1034ymsigg2a-western-blot-human-brain-tissue-lysate-20-g.jpg)
Western blot - Anti-GFAP antibody [EPR1034Y] (ab279290)All lanes : Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (ab279290) at 1/1000 dilution
Lane 1 : Human brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Observed band size: 40-50 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with the literature (PMID: 824020, 2294, 6340792).
Exposure times: Lane 1: 3.25 seconds; Lane 2: 48 seconds; Lane 3: 10 seconds.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab279290)](https://www.abcam.com/ps/products/279/ab279290/Images/ab279290-450188-anti-gfap-antibody-EPR1034Y-immunohistochemistry-cerebral-cortex-human-YR061518PS-MsIgG2a.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab279290)IHC image of GFAP staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab279290, 1µg/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG2a, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit compact polymer system. DAB was used as the chromogen.
The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Immunohistochemistry (Frozen sections) - Anti-GFAP antibody (ab279290)Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling GFAP with ab279290 at 1/500 (1.634 µg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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Flow Cytometry (Intracellular) - Anti-GFAP antibody (ab279290)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized rat primary neural/glia cells labelling GFAP with ab279290 at 1/1000 dilution (0.1µg)/ Right compared with a Mouse monoclonal IgG isotype control/ Left. Goat Anti-Mouse IgG (Alexa Fluor® 647, ab150119) at 1/2000 dilution was used as the secondary antibody.
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![Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (ab279290)](https://www.abcam.com/ps/products/279/ab279290/Images/ab279290-450223-anti-gfap-antibody-epr1034ymsigg2a-immunoprecipitation-rat-brain-tissue-lysate10-ug.jpg)
Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (ab279290)GFAP was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 µg with ab279290 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279290 at 1/1000 dilution. mouse IgG for IP (HRP) (ab131368) was used at 1/5000 dilution.
Lane 1: Rat brain tissue lysate 10µg.
Lane 2: ab279290 IP in rat brain tissue lysate.
Lane 3: Mouse monoclonal IgG2a (ab18413) instead of ab279290 in rat brain tissue lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 23 seconds.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab279290 has not yet been referenced specifically in any publications.