Mouse Monoclonal Antibody to GFAP (GFAP) Cat# MCA-3E10

      Glial fibrillary acidic protein (GFAP) is strongly and specifically expressed in astrocytes, Bergmann glia, certain other glia in the central nervous system, in satellite cells in peripheral ganglia, and in non-myelinating Schwann cells in peripheral nerves. GFAP expression is also seen in developing neural stem cells and GFAP levels may greatly increase in regions of CNS injury or disease. The formation of a GFAP rich "glial scar" following CNS injury may be one reason why reconnection of severed processes is relatively inefficient in adults. Point mutations in the GFAP gene are causative of Alexander disease (5). All forms of Alexander disease are characterized by the presence of Rosenthal fibers, which are GFAP containing cytoplasmic inclusions found in astrocytes. Some interest has recently been focused on GFAP as a protein released into blood and CSF following traumatic brain injury, stroke and other CNS compromises (6,7). Measurement of the levels of blood or CSF GFAP may give information about patient presentation, progress, response to therapy or outcome.

This antibody was made against recombinant human GFAP and works well on human tissues but is less efficient on rodent material. The epitope for this antibody has been mapped to the N-terminal region of the α-helical coiled-coil "rod" region of human GFAP isotype I, and further studies show that the human peptide EIRFLRKIHEEEVRE (196-210) strongly inhibits binding of the antibody to human GFAP. This peptide is conserved between pig and human GFAP, but that of rat and mouse are both slightly divergent, having a glutamine instead of the arginine (see outlined in red below). This small divergence is likely the reason for the lower efficiency of this antibody on rodent material.

Human   EIRFLRKIHEEEVRE (196-210)
Rat     EIQFLRKIHEEEVRE (194-208)
Mouse   EIQFLRKIYEEEVRE (193-207)




Chromogenic immunostaining of a formalin fixed paraffin embedded human brain cortex section with mouse mAb to GFAP, MCA-3E10, dilution 1:1,000, detected with DAB (brown) using the the Vector Labs ImmPRESS method and reagents with citrate buffer retrieval. Hematoxylin (blue) was used as the counterstain. In cortex, GFAP strongly labels astrocytes and glial cells. This antibody performs well in staining with 4% PFA or standard NBF fixed rat and human tissues. MCA-3E10 is our recommended and preferred clone for GFAP paraffin staining applications. Mouse select image for larger view.



Immunofluorescent analysis of mouse hippocampus section stained with mouse mAb to GFAP, MCA-3E10, dilution 1:500 in green, and costained with chicken pAb to NF-L, CPCA-NF-L, dilution 1:2,000, in red. The blue is Hoechst staining of nuclear DNA. Following transcardial perfusion of mouse with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45μM, and free-floating sections were stained with the above antibodies. The GFAP antibody stains network of glial cells while the CPCA-NF-L antibody labels axons and dendrites of neuronal cells. While the staining is convincing it is not as strong as seen with other GFAP antibodies, likely due to epitope issue descussed above. So for rodent studies with GFAP antibodies we recommend an alternate mouse monoclonal MCA-5C10 or one of our polyclonal reagents made in rabbit, RPCA-GFAP, chicken CPCA-GFAP or goat, GPCA-GFAP. These were all made against recombinant human GFAP and we document that they work equally well for WB, IF/ICC and IHC on material of human, rodent and other mammalian species origin. Mouse select image for larger view.