Anti-GFAP antibody [EPR1034Y] (ab279290)
![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 "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
- Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-GFAP antibody [EPR1034Y]
See all GFAP primary antibodies -
Description
Mouse monoclonal [EPR1034Y] to GFAP -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt (Intra) RatIHC-Fr MouseIHC-P HumanIP RatWB MouseRatHuman
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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: Rat primary neural/glia cells. IP: Rat brain tissue lysate.
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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
Images
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Western blot - Anti-GFAP antibody [EPR1034Y] (ab279290)All lanes : Anti-GFAP antibody [EPR1034Y] (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.
