GFAP Polyclonal antibody

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• KD/KO Validated

GFAP Polyclonal Antibody for IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IP, IHC, IF, ELISA

Conjugate

Unconjugated

Publications

4

Cat no : 23935-1-AP

Synonyms

GFAP

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Validation Data Gallery

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  WB analysis of mouse brain using 23935-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 23935-1-AP (GFAP antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse brain using 23935-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain slide using 23935-1-AP (GFAP Antibody) at dilution of 1:50.

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  IHC staining of mouse brain using 23935-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain slide using 23935-1-AP (GFAP Antibody) at dilution of 1:50.

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  IF Staining of mouse brain using 23935-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 23935-1-AP (GFAP antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  WB analysis of U-251 using 23935-1-AP

  WB result of GFAP antibody (23935-1-AP; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-GFAP transfected U-251 cells.

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  IP experiment of mouse brain using 23935-1-AP

  IP Result of anti-GFAP (IP:23935-1-AP, 4ug; Detection:23935-1-AP 1:1000) with mouse brain tissue lysate 4000ug.

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Tested Applications

• Positive WB detected in: mouse brain tissue, C6 cells, U-251 cells
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: mouse brain tissue, human gliomas tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: mouse brain tissue

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:500-1:2000 for WB
• Immunohistochemistry (IHC): IHC : 1:20-1:200
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Published Applications

• WB: See 1 publications below
• IF: See 3 publications below

Product Information

The immunogen of 23935-1-AP is GFAP Fusion Protein expressed in E. coli.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: GFAP fusion protein Ag20853
• Full Name: glial fibrillary acidic protein
• Calculated molecular weight: 432 aa, 50 kDa
• Observed molecular weight: 45-50 kDa
• GenBank accession number: BC013596
• Gene symbol: GFAP
• Gene ID (NCBI): 2670
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

Function

GFAP (Glial fibrillary acidic protein) is a type III intermediate filament (IF) protein specific to the central  nervous system (CNS). GFAP is one of the  main components of the intermediate filament network in astrocytes and has been proposed as playing a role in cell migration, cell motility, maintaining mechanical strength, and in mitosis.

Tissue specificity

GFAP is expressed in central nervous system cells, predominantly in astrocytes. GFAP is commonly used as an astrocyte marker.  However, GFAP is also present in peripheral glia and in non-CNS cells, including fibroblasts, chondrocytes, lymphocytes, and liver stellate cells (PMID: 21219963).

Involvement in disease

• Mutations in GFAP lead to Alexander disease (OMIM: 203450), an autosomal dominant CNS disorder. The mutations  present in affected individuals are thought to be gain-of-function. 

• Upregulation of GFAP is a hallmark of reactive astrocytes, in which GFAP is present in hypertrophic cellular  processes. Reactive astrogliosis is present in many neurological disorders, such as stroke, various  neurodegenerative diseases (including Alzheimer's and Parkinson's disease), and neurotrauma. 

Isoforms

Astrocytes express 10 different isoforms of GFAP that differ in the rod and tail domains (PMID: 25726916),  which means that they differ in molecular size. Isoform expression varies during the development and across different subtypes of astrocytes. Not all isoforms are upregulated in reactive astrocytes.

Post-translational modifications

Intermediate filament proteins are regulated by phosphorylation. Six phosphorylation sites have been identified in GFAP protein, at least some of which are reported to control filament assembly (PMID: 21219963).

Cellular localization

GFAP localizes to intermediate filaments and stains well in astrocyte cellular processes.

Publications

Species	Application	Title
	IF	Sci Rep; Label-free classification of neurons and glia in neural stem cell cultures using a hyperspectral imaging microscopy combined with machine learning.; Authors - Hiroshi Ogi
human	WB	Int J Oncol; Effects of three-dimensional collagen scaffolds on the expression profiles and biological functions of glioma cells.; Authors - Wei Jia
mouse	IF	Biochem Biophys Res Commun; Identification and immunohistochemical characterization of G-quadruplexes in mouse brain.; Authors - Sefan Asamitsu
mouse	IF	Sci Rep; Administration of molecular hydrogen during pregnancy improves behavioral abnormalities of offspring in a maternal immune activation model.; Authors - Kenji Imai