Protein Overview
GFAP is the principal class III intermediate filament protein of mature astrocytes in the CNS, forming the astrocytic cytoskeleton and supporting cell shape, process extension, blood-brain barrier integrity, and synaptic modulation. GFAP is markedly upregulated during reactive astrogliosis following CNS injury or disease.
Expected Localization
Cytoplasmic, filamentous - forms a fibrillar meshwork throughout the astrocyte cell body and processes, including perivascular end-feet.
Immunofluorescence typically shows a stellate, web-like cytoplasmic filament pattern outlining astrocyte morphology.
Expected Expression Pattern
Expressed specifically in astrocytes and some ependymal/radial glial cells of the CNS, with lower levels in enteric glia and non-myelinating Schwann cells. Expression increases substantially with reactive gliosis (injury, neurodegeneration, glioma) and varies with CNS region and disease state.
Relevant Pathways
- Astrocyte cytoskeletal organization / intermediate filament dynamics
- Reactive astrogliosis and neuroinflammatory signaling
- Blood-brain barrier maintenance
- Glial scar formation after CNS injury
- Neurodegenerative disease-associated astrocyte activation
- Glioma/astrocytoma tumor biology
Clone Information
Antibody Species: Mouse
Cell Type Compatibility: Validated in human cells only using Cytos™ kits.
For more details on the clone and reactivity with other species, visit ABclonal and search for the clone catalog # A28062BCS.
Qualitative Results
Localization Assessment
GFAP protein panel demonstrates strong localization to intermediate filaments in GFAP-positive cell line. The observed localization of GFAP protein is consistent with published data.
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| U251 (astrocyte) | HMC3 (microglia) |
U251 and HMC3 cells were cultured in a 12-well format Cytos slide kit overnight then fixed with 4% formaldehyde for 30 minutes, permeabilized using 70% ethanol for 10 minutes. The slide kit was then assembled into a flow cell and loaded onto an AVITI24™ along with the Cytos Protein Cartridge for a multiomics cell screening run. GFAP demonstrated positive expression in U251 cells (astrocyte) and negative expression in HMC3 (microglia), and high signal-to-noise ratio, with no detectable probe crosstalk. These results support high confidence in target-specific detection and validate this antibody as a passing clone.
Quantitative Results
| Signal-to-noise ratio Acceptance criteria: > 2x background |
Pass |
- 37.0 ± 1.6 (U251, GFAP-positive; n=2 runs)
Indicates the fold brightness of the target signal as measured by the Mean Intensity Above (Otsu-defined) Threshold (IAT), over the background (a channel-matched isotype control) in the same cell. Data shown are mean ± standard deviation across independent reproducibility runs. The result of 37.02 in the GFAP-positive U251 line is over 18 times the acceptance threshold — a strong, easily distinguishable signal.
|
Panel-level run-to-run reproducibility |
Pass |
For each imaging panel, we assess reproducibility by comparing the intensity signal a given antibody target produces across independent replicate runs. Within each run, we measure the average per-well fluorescence intensity for every target in the panel. We then plot each target's intensity from one run against the matching target's intensity from another run of the same layout, and calculate the Pearson correlation coefficient (r) between those two sets of measurements. Squaring that value gives the R² (coefficient of determination), which reports what fraction of the variation in one run's signal is explained by the other run's signal.
Disclaimer: This information is provided to help assess whether the target suits your experiment. It reflects internal testing conditions and does not guarantee performance in your application. Performance in other cell types and species such as rat or rabbit may require additional validation using Cytos kits as performance is not guaranteed.
ELEMENT BIOSCIENCES For Research Use Only. Not for Use in diagnostic procedures.