概述

  • 产品名称
  • 描述
    兔多克隆抗体to GFAP
  • 特异性
    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.
  • 经测试应用
    适用于: IHC-FoFr, IHC-Fr, ICC/IF, WB, IHC-P, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Cat, Dog, Human, Common marmoset
    预测可用于: Cow, Pig, Mammal
  • 免疫原

    Full length native protein (purified) corresponding to Human GFAP.

  • 阳性对照
    • IHC-P: FFPE mouse brain normal. IHC-P: FFPE rat brain normal.
  • 常规说明
    In some cases, the antibody may appear red in color. This is due to small amounts of hemolysis, and does not affect antibody performance.

性能

应用

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.

应用 Ab评论 说明
IHC-FoFr 1/5000. See Abreview.
IHC-Fr 1/500.
ICC/IF 1/1000.
WB 1/10000. Detects a band of approximately 55,48 kDa.

This lower 48kDa band is thought to be a degradation product.

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.

靶标

  • 功能
    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.
  • 组织特异性
    Expressed in cells lacking fibronectin.
  • 疾病相关
    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.
  • 序列相似性
    Belongs to the intermediate filament family.
  • 翻译后修饰
    Phosphorylated by PKN1.
  • 细胞定位
    Cytoplasm. Associated with intermediate filaments.
  • Information by UniProt
  • 数据库链接
  • 别名
    • wu:fb34h11 antibody
    • ALXDRD antibody
    • cb345 antibody
    • etID36982.3 antibody
    • FLJ42474 antibody
    • FLJ45472 antibody
    • GFAP antibody
    • GFAP_HUMAN antibody
    • gfapl antibody
    • Glial fibrillary acidic protein antibody
    • Intermediate filament protein antibody
    • wu:fk42c12 antibody
    • xx:af506734 antibody
    • zgc:110485 antibody
    see all

图片

  • Immunohistochemistical 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.

    See Abreview

  • Immunocytochemistry/Immunofluorescence of mixed neuron-glial cultures labelling rabbit GFAP (red channel) and chicken vimentin CPCA-Vim (green channel). The fibroblastic cells contain only vimentin and so are green, while astrocytes contain either vimentin and GFAP, so appearing golden, or predominantly GFAP, in which case they appear red. Blue is nuclear DNA stain.

  • 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 (pH6, 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 haematoxylin 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.

  • 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 (pH6, 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 haematoxylin 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.

  • 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 ~55kDa corresponds to rodent GFAP. A lower band at ~48kDa is derived from the GFAP molecule.

  • All lanes : Anti-GFAP antibody (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 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

  • 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. ab96883 was used as the secondary antibody at a dilution of 1/200.

    See Abreview

  • IHC - Wholemount of rat retina tissue labelling GFAP (red) with ab7260. Sample was incubated with primary antibody (1/100) for 18 hours at 4°C. A Phycoerythrin-conjugated goat anti-rabbit IgG monoclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • 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.

    See Abreview

  • ab7260 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.

    See Abreview

  • Anti-GFAP antibody (ab7260) at 1/5000 dilution + Rat Brain
  • Immunohistochemical analysis of formaldehyde-fixed paraffin-embedded canine neuronal tissue sections, labelling GFAP with ab7260 at a dilution of 1/1000 incubated for 90 minutes at 25°C. Antigen retrival was with 10mM citrate pH6.0 (heat mediated). Blocking was with 10% serum incubated for 30 minutes at 25°C. Secondary was a Goat anti-rabbit polyclonal Texas Red® conjugate at 1/200.

    See Abreview

  •  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 visualised using DAPI.

    See Abreview

  •  ab7260 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.

    See 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.

    See Abreview

文献

This product has been referenced in:
  • Zhao B  et al. Hyperbaric oxygen attenuates neuropathic pain and reverses inflammatory signaling likely via the Kindlin-1/Wnt-10a signaling pathway in the chronic pain injury model in rats. J Headache Pain 18:1 (2017). IHC ; Rat . Read more (PubMed: 28058534) »
  • Kim HJ & Han SJ A simple rat model of mild traumatic brain injury: a device to reproduce anatomical and neurological changes of mild traumatic brain injury. PeerJ 5:e2818 (2017). IHC ; Rat . Read more (PubMed: 28070456) »

See all 190 Publications for this product

客户评价及客户问答

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Meriones unguiculatus Tissue sections (Neocortex)
Antigen retrieval step
None
Permeabilization
Yes - Triton X-100
Specification
Neocortex
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Oct 18 2017

Application
IHC - Wholemount
Sample
Mouse Tissue (Intestine)
Specification
Intestine
Username

Abcam user community

Verified customer

提交于 Oct 12 2017

Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Brain, astrocytes)
Specification
Brain, astrocytes
Username

Abcam user community

Verified customer

提交于 Jul 14 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Substantia nigra, astrocytes)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Substantia nigra, astrocytes
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Edward Fletcher

Verified customer

提交于 Jul 13 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, PH6.0
Permeabilization
Yes - 0.1% TritonX-100
Specification
Brain
Blocking step
Cas-Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 30 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (sciatic nerve)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization
No
Specification
sciatic nerve
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 25 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Tris-Acetate)
Loading amount
20 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Mar 15 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Guinea pig Tissue lysate - whole (Hippocampus)
Gel Running Conditions
Reduced Denaturing (7,5)
Loading amount
10 µg
Specification
Hippocampus
Blocking step
ecl prime blocking reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Feb 24 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: CC1
Permeabilization
No
Specification
Brain
Fixative
Formalin
Username

Dr. Emmanuel Bouchaert

Verified customer

提交于 Jan 03 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cell (brain)
Permeabilization
Yes - 4%paraform
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde
Username

Kyungjoo Seong

Verified customer

提交于 Nov 29 2016

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