概述

  • 产品名称Anti-GFAP抗体[GF5]
    参阅全部 GFAP 一抗
  • 描述
    小鼠单克隆抗体[GF5] to GFAP
  • 特异性There is no cross-reactivity with other neurospecific proteins.
  • 经测试应用适用于: IHC-P, WB, ELISA, IHC-FoFr, IHC-Fr, ICC/IF, Flow Cyt, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    This clone has been derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunised with purified glial fibrillary acidic protein from human brain.

  • 阳性对照
    • normal adult rat brain: lateral ventricle IHC-P: FFPE human hippocampus normal. IHC-P: FFPE rat brain normal.
  • 常规说明Concentration varies from lot to lot and can be provided on request.

性能

应用

Our Abpromise guarantee covers the use of ab10062 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-P 1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 43-45 kDa.
ELISA Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 20708681
IHC-Fr 1/1000.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1-2µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC 1/100.

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

Anti-GFAP antibody [GF5] 图像

  • IHC image of GFAP staining in a formalin fixed, paraffin embedded rat normal 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 ab10062 at 1/100 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.

  • IHC image of GFAP staining in a formalin fixed, paraffin embedded human normal hippocampus 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 ab10062 at 1/500 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.

     

  • Overlay histogram showing SH-SY5Y cells stained with ab10062 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab10062, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Mouse monoclonal to GFAP [GF5] (ab10062) was used in fixed murine cultures (mixed neurons/glia) at 1/100 overnight at 4°C. A secondary goat anti-mouse antibody was used for detection (Alexa Fluor 568; 1/400). Microscopy revealed diffuse cytosolic labelling. Coounterstaining with TO-PRO-3 (Molecular Probes; 660nm (converted here to blue colour) was used to identify the nucleus. The “fibrous” anti-GFAP staining of murine mixed cultures is typical of what is expected.

  • GFAP antibody [GF5] - Astrocyte Marker (ab10062) immunocytochemical detection in stimulated Cor1 cells. Stimulated Cor1 cells were fixed in formaldehyde, permeabilized, blocked in 1% BSA for 10 mins @ rt°C. Primary Antibody ab10062 incubated at 1/1500 for 2 hours in TBS/BSA/azide/0.3% triton. Secondary Antibody: anti mouse IgG Conjugated to: Alexa Fluor® 488 (1/1000).

    See Abreview

  • GFAP antibody [GF5] - Astrocyte Marker (ab10062; 1/250 for 16h) used in Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) on Rat Tissue sections (adult brain: lateral ventricle showing astrocytes).Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Secondary Antibody: biotin labelled goat anti mouse Ig (1/200). This image shows the exit point for the progenitor olfactory neurones, of the lateral ventrical subventricular zone.
  • Mouse monoclonal to GFAP [GF5] (ab10062) was used in fixed rat glial cultures at a dilution of 1/100, and incubated overnight at 4°C. Alexa Fluor 568 (1/400) secondary goat anti-mouse antibody was used for detection. Fluoresence microscopy revealed diffuse cytosolic labelling. Counterstaining with TO-PRO-3 (Molecular Probes; 660nm (converted here to blue colour) was used to identify the nucleus. The “fibrous” anti-GFAP staining of murine mixed cultures is typical of what is expected.

Anti-GFAP antibody [GF5] (ab10062)参考文献

This product has been referenced in:
  • Zhai W  et al. A1 adenosine receptor attenuates intracerebral hemorrhage-induced secondary brain injury in rats by activating the P38-MAPKAP2-Hsp27 pathway. Mol Brain 9:66 (2016). Rat . Read more (PubMed: 27301321) »
  • Murlidharan G  et al. Unique glycan signatures regulate adeno-associated virus tropism in the developing brain. J Virol 89:3976-87 (2015). ICC/IF ; Mouse . Read more (PubMed: 25631075) »

See all 20 Publications for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (spinal cord cross section, 25 micrometer thickness)
Permeabilization No
Specification spinal cord cross section, 25 micrometer thickness
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative Formaldehyde
Username

Mr. Tamas Bellak

Verified customer

提交于 Nov 25 2015

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Mouse Tissue sections (frozen brain section)
Specification frozen brain section
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 23 2013

Application Flow Cytometry
Fixation Frozen
Permeabilization Yes - 0.1% Triton-X
Sample Human Cell (Brain)
Specification Brain
Gating Strategy forward scatter/side scatter (all cells)
Preparation Cell harvesting/tissue preparation method: Homogenized frozen tissue
Sample buffer: Tris/EDTA/sucrose/Triton
Username

Dr. Mollie Woodworth

Verified customer

提交于 Sep 03 2013

Thank you for contacting us. I apologise for the delay getting back to you.

Several markers can be used to stain astrocytes: alpha Smooth Muscle Actin, Bystin, GFAP, GLT-1, GLAST, S100, Mab 6.17, SC1, etc…

I have contacted my...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - 0.03% Trition
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Astrocytes)
Loading amount 30 µg
Specification Astrocytes
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 08 2013

Thank you for getting back to me.

I could certainly arrange the new vial (ab7260) as a replacement for you as requested.

However, after reading through all the previous correspondence, I would like to highlight and advise the follow...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Thank you for getting back to me. I am very sorry to hear that the product does not perform.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means th...

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Thank you for your enquiry regarding ab10062 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

This antibody has not been tested specifically in...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"