Invitrogen

GFAP Monoclonal Antibody (ASTRO6)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated. View Details

6 Published Figures

46 References

View all (81) GFAP antibodies

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Datasheet

Questions & Answers

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GFAP Antibody in Western Blot (WB) — Western blot was performed using Anti-GFAP Monoclonal Antibody (ASTRO6), (Product # MA5-12023) and a 50 kDa band corresponding to GFAP was observed in the tissues tested along with Mouse IgG at ~25 kDa. Whole cell extracts (30 µg lysate) of Mouse brain (Lane 1), Rat brain (Lane 2), Mouse liver (Lane 3), and Rat liver (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.2 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

GFAP Antibody in Immunohistochemistry (IHC) — Formalin-fixed, paraffin-embedded human brain stained with GFAP antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of astrocytes.

GFAP Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of GFAP showing staining in the cytoplasm of paraffin-embedded rat brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a GFAP monoclonal antibody (Product # MA5-12023) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

GFAP Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Immunofluorescent analysis of beta-3 Tubulin (green) and GFAP (red) in human iPSC-derived forebrain organoids derived at Day 84. The organoids were fixed with 4% PFA for 1 hour at room temperature, followed by incubation with 30% sucrose solution overnight at 4°C. The organoids were then embedded in OCT and cryosectioned at 5 µm, permeabilized with 0.2% Triton X-100 for 20 min, and blocked with 10% donkey serum in PBS for 30 min at room temperature. Organoid slices were stained with a Mouse beta-3 Tubulin monoclonal antibody (green; Product # MA1-118) at a dilution of 1:500, and a Rat GFAP monoclonal antibody (red; Product # 13-0300) at a dilution of 1:500 in blocking buffer overnight at 4°C, and then incubated with Donkey anti-Mouse Alexa Fluor 488 (Product # R37114) and Donkey anti-Rat Alexa Fluor 568 (Product # A-11077) at a dilution of 1:1000 as well as DAPI (blue; 1:25000) in blocking solution at room temperature for 1 hour. Images were takenat 20X magnification. Data courtesy of Dr. Zhexing Wen at Emory University.

GFAP Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of glial fibrillary acidic protein (GFAP) in E18 Sparague Dawley primary cortical neuronal cells containing astrocytes. The cells were fixed with 4% formaldehyde for 15 mins, permeabilized with 0.25% Triton X-100 in PBS for 10 mins, and blocked with 3% BSA in PBS for 30 mins at RT. Cells were stained with a GFAP mouse monoclonal antibody (Product # MA5-12023) at a dilution of 1:200 in 3% BSA in PBS for 1 hr at RT, and then incubated with Invitrogen™ AlexaFluor™ 488 Plus goat anti-mouse IgG secondary antibody (Product # A32723) at a dilution of 1:1000 for 1 hr at RT. Nuclei were stained with Hoechst 33342 (Product # H3570). The image contains overlay of GFAP (green) and nuclei (blue). Images were taken on a Zeiss LSM 710 confocal microscope at 40X magnification.

GFAP Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissue tested owing to their inherent genetic constitution. Relative expression of GFAP was observed in Mouse brain, Rat brain in comparison to Mouse kidney and Rat kidney using Anti-GFAP monoclonal antibody (ASTRO6) (Product # MA5-12023) in Western Blot. (doi: 10.1016/j.tins.2015.04.003). {RE}

GFAP Antibody — Fig 4 Histological analysis performed on paraffin-embedded slices of resected GB of a rat. A. H&E staining confirmed the presence of GB characterized by central tumor necrosis (1), tumor (2) and the peritumoral zone with infiltrating cancer cells (3) surrounded by healthy brain tissue (4). B. Immunohistochemistry for Ki67 indicated that GB is highly proliferative, except for the necrotic tumor core: tumor necrosis (1), tumor (2), peritumoral zone with infiltrating cancer cells (3) and healthy brain tissue (4). C-D. Immunohistochemistry for GFAP and Cx43 demonstrated GFAP-positive reactive astrocytes and enhanced Cx43 expression at the peritumoral zone.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1-2 µg/mL

View 1 publication 1 publication

Immunohistochemistry (IHC)

View 25 publications 25 publications

Immunohistochemistry (Paraffin) (IHC (P))

1-2 µg/mL

View 4 publications 4 publications

Immunohistochemistry (Frozen) (IHC (F))

Assay Dependent

View 3 publications 3 publications

Immunocytochemistry (ICC)

View 9 publications 9 publications

Immunofluorescence (IF)

View 3 publications 3 publications

Flow Cytometry (Flow)

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1:20-1:200

Product Specifications

Species Reactivity

Chicken, Human, Pig, Rat

Published species

Human, Mouse, Pig, Rat

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

ASTRO6

Immunogen

Glial Fibrillary Acidic Protein

Conjugate

Unconjugated

Form

Liquid

Concentration

0.2 mg/mL

Purification

Protein G

Storage buffer

PBS, pH 7.4, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10984338

Product Specific Information

MA5-12023 targets Glial Fibrillary Acidic Protein in IF, IHC (P), and WB applications and shows reactivity with Chicken, Human, Porcine, and Rat samples.

The MA5-12023 immunogen is glial Fibrillary Acidic Protein.

MA5-12023 was successfully used to detect astrocytes in E18 Sparague Dawley primary cortical cells.

Target Information

GFAP (Glial fibrillary acidic protein) is a member of the class III intermediate filament protein family. GFAP is heavily and specifically expressed in astrocytes and certain astroglia of the central nervous system, in satellite cells of peripheral ganglia, and in non-myelinating Schwann cells of peripheral nerves. In addition, neural stem cells strongly express GFAP. Antibodies to GFAP are very useful as markers of astrocytic cells. In addition, many types of brain tumor, presumably derived from astrocytic cells, heavily express GFAP. GFAP is also found in the lens epithelium, Kupffer cells of the liver, in some cells in salivary tumors and has been reported in erythrocytes. GFAP is used as a marker to distinguish astrocytes from other glial cells during development. Mutations in this gene cause Alexander disease, a rare disorder of astrocytes in the central nervous system. Alternative splicing of the GFAP gene results in multiple transcript variants encoding distinct isoforms.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.