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.

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.

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)

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.

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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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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 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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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.