Anti-GFAP 抗体 (ab7260)
Key features and details
- Rabbit polyclonal to GFAP
- Suitable for: IHC (PFA fixed), WB, IHC-P, ICC
- Reacts with: Mouse, Rat
- Isotype: IgG
製品の概要
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製品名
Anti-GFAP antibody
GFAP 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to GFAP -
由来種
Rabbit -
特異性
Specifically recognizes mammalian GFAP on western blots and immunocytochemically. Detects a band of 55kDa corresponding to GFAP and also a GFAP derived 48kDa 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. -
アプリケーション
適用あり: IHC (PFA fixed), WB, IHC-P, ICCmore details -
種交差性
交差種: Mouse, Rat
交差が予測される動物種: Horse, Cow, Human, Pig, Mammals -
免疫原
Recombinant full length protein corresponding to Human GFAP. Isotype 1 expressed in and purified from E. coli.
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特記事項
In some cases, the antibody may appear red in color. This is due to small amounts of hemolysis, and does not affect antibody performance.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
Preservative: 0.01% Sodium azide -
Concentration information loading...
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精製度
Whole antiserum -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
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.
アプリケーション | Abreviews | 特記事項 |
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IHC (PFA fixed) | Use at an assay dependent concentration. | |
WB | 1/10000. Detects a band of approximately 55,48 kDa. This lower 48kDa band is thought to be a degradation product. |
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IHC-P | 1/1000 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
ICC | 1/5000. |
ターゲット情報
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機能
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. -
組織特異性
Expressed in cells lacking fibronectin. -
関連疾患
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. -
配列類似性
Belongs to the intermediate filament family. -
翻訳後修飾
Phosphorylated by PKN1. -
細胞内局在
Cytoplasm. Associated with intermediate filaments. - Information by UniProt
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参照データベース
- 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 -
別名
- wu:fb34h11 antibody
- ALXDRD antibody
- cb345 antibody
see all
画像
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GFAP antibody ab7260 was used with Tissue Clearing Kit ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain. Blue: DAPI, Green: GFAP.
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.
To use this antibody with tissue clearing, use Tissue Clearing Kit ab243298. For 1 mm brain sections, we recommend a starting dilution of 1:1000, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (ab150077) at a dilution of 1:400.
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All lanes : Anti-GFAP antibody (ab7260) at 1/5000 dilution
Lanes 1-3 : Rat thoracotomy, spinal cord homogenate
Lanes 4-5 : Rat thoracotomy sham, spinal cord homogenate
Lanes 6-7 : Rat nerve transect sham, spinal cord homogenate
Lanes 8-9 : Rat nerve transect, spinal cord homogenate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : 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
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IHC image of GFAP staining in a formalin fixed, paraffin embedded normal mouse brain tissue section, performed on a Leica Bond™ system using the standard protocol B.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7260 at 1/2000 dilution for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)Yoon, H. et al PLoS One. 2017 Jul 10;12(7):e0180697. doi: 10.1371/journal.pone.0180697. eCollection 2017 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Increases in GFAP after demyelinating injury are greater in the spinal cord compared to brain
Photomicrographs show immunoreactivity for GFAP or ALDH1L1 in (A) the corpus callosum, or (B) the dorsal column white matter of adult mice at base line, and at 14 d after microinjection of the demyelinating agent lysolecithin. Histograms show the percent area of GFAP immunofluorescence, and expression of GFAP/ALDH1L1+ astrocyte, was significantly greater in the spinal cord compared to the corpus callosum 14d post-lysolecithin lesion.
GFAP was detected using ab7260.
(From Figure 4A and 4B of Hoon et al)
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All lanes : Anti-GFAP antibody (ab7260) at 1/5000 dilution
Lane 1 : Rat brain lysate
Lane 2 : Rat spinal cord lysate
Lane 3 : Mouse brain lysate
Lane 4 : Mouse spinal cord lysate -
IHC image of GFAP staining in a formalin fixed, paraffin embedded normal rat brain tissue section, performed on a Leica Bond™ system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7260 at 1/2000 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.
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.
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Immunocytochemistry - Anti-GFAP antibody (ab7260)This image is courtesy of an Abreview submitted by Chia-Li Lin.
ab7260 staining GFAP in cells from mouse brain tissue sections by ICC/IF (Immunocytochemistry/immunofluorescence).
Cells were fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 in PBS and blocked with 1% BSA for 40 minutes at 25°C. Samples were incubated with primary antibody (1/1200 in TBS) for 24 hours at 4°C. Goat Anti-Rabbit IgG H&L (DyLight® 488) (ab96883) was used as the secondary antibody at a dilution of 1/200.
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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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)This image is courtesy of an Abreview submitted by Mr Osama Mohsen
ab7260 staining rat brain tissue sections by IHC-P.
Sections were fixed in formaldehyde and blocked with a commercialy available blocking agent prior to incubating with ab7260, diluted 1/5000 for 20 hours at 4°C. An HRP conjugated mouse polyclonal (universal HRP polymer detection) antibody was used as the secondary.
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Immunocytochemistry - Anti-GFAP antibody (ab7260)This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy
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 visualized using DAPI.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)This image was kindly supplied by Dr Vladimir Milenkovic by Abreview
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.
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GFAP antibody ab7260 was used with 3D Cell Culture Clearing Kit ab243299 to penetrate, stain and clear a 3D neuronal spheroid cell culture. Blue: DAPI, Green: GFAP.
Learn more about 3D cell culture clearing and tissue clearing kits, reagents, and protocols designed to make it easier to stain 3D cell cultures and thick tissue sections and get more data from each valuable tissue section.
To use this antibody with clearing, use our 3D Cell Culture Clearing Kit ab243299. We recommend a starting dilution of 1:1000, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (ab150077) at a dilution of 1:400.
プロトコール
参考文献 (557)
ab7260 は 557 報の論文で使用されています。
- Findlay AS et al. Fam151b, the mouse homologue of C.elegans menorin gene, is essential for retinal function. Sci Rep 10:437 (2020). PubMed: 31949211
- Yin N et al. IDH1-R132H mutation radiosensitizes U87MG glioma cells via epigenetic downregulation of TIGAR. Oncol Lett 19:1322-1330 (2020). PubMed: 31966064
- Tu Y et al. Melatonin inhibits Müller cell activation and pro-inflammatory cytokine production via upregulating the MEG3/miR-204/Sirt1 axis in experimental diabetic retinopathy. J Cell Physiol N/A:N/A (2020). PubMed: 32324260
- Li W et al. Dysfunctional Nurr1 promotes high glucose-induced Müller cell activation by up-regulating the NF-?B/NLRP3 inflammasome axis. Neuropeptides N/A:102057 (2020). PubMed: 32461025
- Zhao M et al. Vinpocetine Protects Against Cerebral Ischemia-Reperfusion Injury by Targeting Astrocytic Connexin43 via the PI3K/AKT Signaling Pathway. Front Neurosci 14:223 (2020). PubMed: 32300287