Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)

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.

Carl Hobbs, King`s College London, United Kingdom

Immunocytochemistry/ Immunofluorescence - GFAP antibody - Astrocyte Marker (ab7260)

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.

Immunohistochemistry (Frozen sections) - GFAP antibody - Astrocyte Marker (ab7260)

 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.

This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy

Western blot - GFAP antibody - Astrocyte Marker (ab7260)

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 ~50kDa corresponds to rodent GFAP. A lower band at ~45kDa is derived from the GFAP molecule.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)

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.

This image is courtesy of an Abreview submitted by Mr Osama Mohsen

Western blot - GFAP antibody - Astrocyte Marker (ab7260)

All lanes : Anti-GFAP antibody - Astrocyte Marker (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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody - Astrocyte Marker (ab7260)

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 25ºC; followed by incubation with the primary antibody, at a 1/500 dilution, for 16 hours at 4ºC. 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.

This image was kindly supplied by Dr Vladimir Milenkovic by Abreview

Immunocytochemistry/ Immunofluorescence - GFAP antibody - Astrocyte Marker (ab7260)

 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.

This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy