Anti-GFAP antibody [2A5] - Astrocyte Marker

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [2A5] - Astrocyte Marker (AB4648)

Immunohistochemistry analysis of paraffin-embedded human cerebellum tissue sections labelling GFAP with ab4648 at 1/500 dilution (1 mg/mL). Sections were stained with ab4648 using the HRP/DAB staining. Sections were counterstained with hematoxylin/eosin. Antigen retrieval was heat mediated using Antigen retrieval buffer (100X citrate buffer) (pH 6.0) (ab93678) for 15 minutes.
TTo the right is a region of cerebellar molecular layer containing the prominent cytoskeletal fibers of Bergmann glia which are strongly positive for GFAP. The middle shows a region of the granular layer and to the left is white matter, both of which contain GFAP positive astrocytes. The immunostaining was performed with the Vector ImmPress rat adsorbed horse anti-mouse IgG detection kit.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [2A5] - Astrocyte Marker (AB4648)

ab4648 staining human substantia nigra. Staining is localised to the cytoplasm.
Left panel : with primary antibody diluted 1 : 4000. Right panel : isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [2A5] - Astrocyte Marker (AB4648)

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling GFAP with ab4648 at 0.2 ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab4648 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-FrFl

Supplier Data

Immunohistochemistry - Free Floating - Anti-GFAP antibody [2A5] - Astrocyte Marker (AB4648)

Immunofluorescence analysis of rat cerebellum tissue labeling GFAP with ab4648 at 1/500 dilution (Red), Costained with parvalbumin antibody at 1/2,000 dilution (Green). The blue is DAPI staining of nuclear DNA. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, and free-floating 45μM sections were stained with above antibodies. ab4648 stains the processes of Bergmann glia and astrocytes. The Pvalb antibody labels perikarya and dendrites of Purkinje cells and interneurons in the molecular layer of the cerebellum. The staining on rodent tissues is specific but not as robust as on human material.