Anti-GFAP antibody ab4674 is a chicken polyclonal antibody that is used in GFAP western blotting, IHC and immunofluorescence. Suitable for mouse and rat samples.
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Same trusted quality, new lower price
IgY
Chicken
Preservative: 0.03% Sodium azide
Constituents: 99.97% PBS
Liquid
Polyclonal
IHC-P | ICC/IF | WB | IHC-FrFl | |
---|---|---|---|---|
Human | Tested | Expected | Expected | Expected |
Mouse | Tested | Expected | Expected | Tested |
Rat | Tested | Tested | Tested | Expected |
Mammals | Predicted | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/200 - 1/20000 | Notes - |
Species Mouse | Dilution info 1/200 - 1/20000 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/200 - 1/20000 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mammals | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/500 - 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mammals | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/1000 - 1/5000 | Notes Expect to see a band at 55kDa and another at about 48kDa, apparently a breakdown product of the 55kDa band. |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mammals | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/5000 | Notes Try this antibody at about between about 1:1,000 using fluorescent secondary antibodies or 1:5,000 using peroxidase or other enzyme linked methods. |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mammals | Dilution info - | Notes - |
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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.
Glial fibrillary acidic protein, GFAP, GFAP
Anti-GFAP antibody ab4674 is a chicken polyclonal antibody that is used in GFAP western blotting, IHC and immunofluorescence. Suitable for mouse and rat samples.
- Tried and trusted by researchers since 2003
Same trusted quality, new lower price
IgY
Chicken
Preservative: 0.03% Sodium azide
Constituents: 99.97% PBS
Liquid
Polyclonal
IgY fraction
Concentrated IgY fraction of egg yolks.
Blue Ice
+4°C
Do Not Freeze
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This supplementary information is collated from multiple sources and compiled automatically.
GFAP or Glial Fibrillary Acidic Protein plays a critical role in the structure and function of astrocytes which are star-shaped glial cells in the brain and spinal cord. It is a type III intermediate filament protein with a molecular weight of approximately 50 kDa. GFAP mainly expresses in the central nervous system specifically within astrocytes but you can also find it in non-neuronal cells such as enteric glia. Researchers frequently use GFAP antibodies in laboratory techniques like GFAP western blot and GFAP IHC to study protein expression and localization.
GFAP contributes to the structural integrity and functional maintenance of the cytoskeleton in astrocytes. This protein integrates into a network of intermediate filaments providing mechanical support to cells. The ability of GFAP to form homodimers and filamentous structures is vital for its function. GFAP antibodies help in highlighting these structures in studies and techniques such as GFAP ELISA utilize these antibodies to quantify protein levels. Through GFAP staining scientists gain insights into the cellular architecture and any alterations under pathological conditions.
GFAP plays a significant role within the neurological framework of cell signaling and response to injury. It links to the MAPK signaling pathway which modulates cell growth and differentiation. This protein also interacts with vimentin another intermediate filament protein forming a complex that affects astrocytic responses to environmental changes. These interconnected pathways and interactions highlight GFAP's importance in maintaining homeostasis within the central nervous system.
GFAP anomalies relate to various neurodegenerative conditions and injuries such as Alexander disease and gliosis. Mutations in the GFAP gene form a significant part of Alexander disease pathology where GFAP accumulates abnormally in astrocytes. Its interaction with vimentin aggravates the disease state affecting the astrocytic function. Researchers utilize anti-GFAP antibodies extensively to study disease progression and potential therapeutic targets providing insights into treatments for conditions involving GFAP dysregulation.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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GFAP immunofluorescence staining of primary hippocampal rat neurons/glia using chicken anti-GFAP antibody
ab4674 staining GFAP in primary hippocampal rat neurons/glia (obtained from Neuromics cat. no. PC35101) DIV14. The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab4674 at 1μg/ml and Anti-Aldolase C antibody - Astrocyte Marker ab87122 Rabbit Poly to Mouse Fructose-bisphosphate aldolase C (No Modifications). Cells were then incubated with Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat polyclonal Secondary Antibody to Chicken IgY - H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution (shown in red) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed at 1/1000 dilution (shown in pseudocolour green). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
GFAP immunofluorescence staining of primary mouse neurons/glia using chicken anti-GFAP antibody
ab4674 staining GFAP in primary mouse neurons/glia DIV14 (prepared from E18 mouse hippocampal brain area obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) cells. The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab4674 at 1µg/ml and Anti-L1CAM antibody [2C2] ab24345 Mouse mono Anti-L1CAM [2C2]. Cells were then incubated with Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat polyclonal Secondary Antibody to Chicken IgY - H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
GFAP antibody ab4674 was used with Tissue Clearing Kit Tissue Clearing Kit - hydrophobic ab243298 to penetrate, stain and clear a 500 μm section of rat brain.
Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain thick tissue sections and get more data from each valuable tissue section.
10x and 63x z-stack images of the CA1 hippocampal region for each treatment are shown, with NeuN staining in green and GFAP staining in red.
Two month-old wild-type mice were placed on either control (n = 10) or inhibitor diet (PLX3397, provided at 290 mg/kg chow; n = 14) for 21 d, causing the elimination of approximately 99% of microglia brain-wide.
Fluorescent immunolabeling of the microglia followed a standard indirect technique (primary antibody followed by fluorescent secondary antibody). Brain tissue (sliced at 40 μm) was stained using the anti-ionized calcium-binding adapter molecule 1 (IBA1, polyclonal, rabbit) antibody (1:1000; Wako, Cat. #019–19741), mounted on slides, and coverslipped using Dapi Fluoromount-G (SouthernBiotech). Half brain images were obtained by stitching using a Zeiss AxioImager M2 upright microscope and Stereo Investigator software package from MicroBrightField. In addition, tissue was stained with anti-hexaribonucleotide binding protein-3 (NeuN, monoclonal, mouse) antibody (1:1000; Millipore; Cat. #MAB377) to label neurons and anti-glial fibrillary acidic protein (GFAP, polyclonal, chicken) antibody (1:500; Abcam; Cat. #ab4674) to label astrocytes, and 10x and 63x z-stack images obtained for each treatment using confocal microscopy.
GFAP immunohistochemistry staining of mouse hippocampus using chicken anti-GFAP antibody
Immunofluorescent analysis of a section of mouse hippocampus stained with ab4674 at a 1:5000 dilution in green.
Costained with a rabbit pAb to FOX3/NeuN dilution 1:5000 in red. The blue is DAPI staining of nuclear DNA. Following transcardial perfusion with 4% paraformaldehyde mouse brain was post fixed for 24 hours cut to 45 μM and free-floating sections were stained. The GFAP antibody stains a network of astroglial cells while the Fox3/NeuN antibody stains the nuclei and proximal perikarya of neurons.
GFAP immunohistochemistry staining of mouse brain using chicken anti-GFAP antibody
IHC image of GFAP staining in a formalin-fixed paraffin-embedded mouse normal brain tissue section.
The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab4674 at 1/1000 dilution for 15 minutes at room temperature. A goat anti-chicken biotinylated secondary antibody was used to detect the primary and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.
All lanes: Western blot - Anti-GFAP antibody (ab4674) at 1/5000 dilution
Lane 1: Rat whole brain lysate
Lane 2: Mouse whole brain lysate
Predicted band size: 49 kDa
GFAP immunohistochemistry staining of rat hippocampus using chicken anti-GFAP antibody
IHC image of GFAP staining in a formalin fixed paraffin embedded normal rat hippocampus tissue section.
The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab4674 at 1/1000 dilution for 15 minutes at room temperature. A goat anti-chicken biotinylated secondary antibody was used to detect the primary and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.
GFAP immunofluorescence staining of primary hippocampal mouse neurons/glia using chicken anti-GFAP antibody
Immunofluorescence staining of SOX9 using Anti-SOX9 antibody [EPR14335] ab185230 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-SOX9 antibody [EPR14335] ab185230 at 5 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and Anti-NeuN antibody [1B7] - Neuronal Marker ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
GFAP immunofluorescence staining of primary hippocampal mouse neurons/glia using chicken anti-GFAP antibody
Immunofluorescence staining of SOX9 using Anti-SOX9 antibody [EPR14335-78] ab185966 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Anti-SOX9 antibody [EPR14335-78] ab185966 at 1 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and Anti-NeuN antibody [1B7] - Neuronal Marker ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative. SOX9 positive cells which are not GFAP positive (e.g. asterisk) are likely neural stem cells/ oligodendrocyte precursor cells present in the culture.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
GFAP immunofluorescence staining of primary hippocampal mouse neurons/glia using chicken anti-GFAP antibody
Immunofluorescence staining of SOX9 using Alexa Fluor® 488 Anti-SOX9 antibody [EPR14335] ab196450 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-SOX9 antibody [EPR14335] ab196450 at 1 μg/ml (shown in green), ab4674 (anti-GFAP) at 1/1000 dilution and Anti-NeuN antibody [1B7] - Neuronal Marker ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
GFAP immunohistochemistry staining of human cerebellum using chicken anti-GFAP antibody
Chromogenic Immunostaining of a formalin fixed paraffin embedded human cerebellum section with chicken pAb to GFAP, dilution 1/20000, detected in DAB (brown) following the ABC method. Hematoxylin (blue) was used as the counterstain. ab4674 detects the core of processes of astrocytes and Bergman glia within the granular and molecular layers. Mouse select image for larger view.
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