| Name: | Goat polyclonal antibody to GFAP |
| Immunogen: | Recombinant full length human GFAP isotype 1 expressed in and purified from E. coli. |
| HGNC Name: | GFAP |
| UniProt: | P14136 |
| Molecular Weight: | ~50kDa |
| Host: | Goat |
| Isotype: | |
| Species Cross-Reactivity: | Human, rat, mouse |
| RRID: | AB_2858264 |
| Format: | Affinity purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3 |
| Applications: | WB, IF/ICC, IHC |
| Recommended Dilutions: | WB: 1:5,000. IF/ICC 1:5,000 |
| Storage: | Store at 4°C for short term, for longer term at -20°C |
Immunofluorescent analysis of mouse hippocampus section stained with goat pAb to GFAP, GPCA-GFAP, dilution 1:5,000 in green, and costained with mouse mAb to FOX3/NeuN, MCA-1B7, 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 above antibodies. The GFAP antibody stains the network of astrocytic glial cells, while the FOX3/NeuN antibody specifically labels nuclei and proximal perikarya of neurons.
Western blot analysis of brain lysates from different species using goat pAb to GFAP, GPCA-GFAP, dilution 1:5,000 in green: [1] protein standard (red), [2] rat cortex, [3] rat cerebellum, [4] mouse cortex, [5] mouse cerebellum, [6] cow cortex, and [7] cow cerebellum. Strong band at about 50 kDa corresponds to GFAP protein. Smaller proteolytic fragments of GFAP are also detected on the blot.
Goat Polyclonal Antibody to GFAP
Cat# GPCA-GFAP
$150.00 – $1,000.00
Glial Fibrillary Acidic Protein (GFAP) is a major CNS protein which runs on SDS-PAGE as a ~50kDa protein, usually associated with somewhat lower molecule weight bands which are alternate transcripts from the single gene or in vivo proteolytic fragments. GFAP is strongly and specifically expressed in astrocytes and certain other glia in the central nervous system, in satellite cells in peripheral ganglia, in non-myelinating Schwann cells in peripheral nerves and is also a useful marker of neural stem cells (1-3). Astrocytes respond to many damage and disease states resulting in “astrogliosis” or the presence of a “glial response”. GFAP antibodies are widely used to study reactive astrocytes which form part of this response, since these cells stain much more strongly with GFAP antibodies than normal astrocytes. GFAP also forms a major component of the so-called glial scar, an astrocyte rich structure apparently forming part of the barrier to nerve fiber regeneration following damage in the central nervous system (4). Neural stem cells frequently strongly express GFAP but many lose this if they develop into neurons or oligodendrocytes. Finally, Alexander disease was recently shown to be caused by point mutations in the protein coding region of the GFAP gene (5). All forms of Alexander disease are characterized by the presence of Rosenthal fibers, which are GFAP containing cytoplasmic inclusions found in astrocytes.
Antibodies to GFAP are therefore very useful as a marker of normal and reactive glial cells in central and peripheral nerve system, as well as of developing neural stem cells.
GPCA-GFAP antibody was made against full length human recombinant GFAP, Prot-r-GFAP, expressed in and purified from E. coli. The antibody works well on western blots, and on immunostaining of cell culture or tissue sections. The same GFAP immunogen was used to produce rabbit, RPCA-GFAP, and chicken, CPCA-GFAP polyclonal antibodies. Using a different immunogens EnCor manufactured a widely used mouse monoclonal antibodies to GFAP, MCA-5C10, MCA-2A5, and MCA-3E10. Mouse select image above left for magnified view.
Chromogenic immunostaining of a formalin fixed paraffin embedded mouse cerebral cortex section with goat pAb to GFAP, GPCA-GFAP, dilution 1:10,000, detected with DAB (brown) using the Vector Elite ABC-HRP detection and reagents with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. In normal brain tissue, the GFAP antibody specifically labels astrocytes. It can also detect chronically activated microglia in neurodegenerative conditions. Mouse select image for larger view.
This antibody was raised against recombinant human GFAP expressed in and purified from E. coli. The human GFAP sequence was derived from NP_002046.1 which was inserted into the eukaryotic expression vector pET30a(+) which adds an N-terminal His tag and some other sequence which is underlined below.
MHHHHHHSSG LVPRGSGMKE TAAAKFERQH MDSPDLGTDD DDKAMADIGS EFMERRRITS 60
AARRSYVSSG EMMVGGLAPG RRLGPGTRLS LARMPPPLPT RVDFSLAGAL NAGFKETRAS 120
ERAEMMELND RFASYIEKVR FLEQQNKALA AELNQLRAKE PTKLADVYQA ELRELRLRLD 180
QLTANSARLE VERDNLAQDL ATVRQKLQDE TNLRLEAENN LAAYRQEADE ATLARLDLER 240
KIESLEEEIR FLRKIHEEEV RELQEQLARQ QVHVELDVAK PDLTAALKEI RTQYEAMASS 300
NMHEAEEWYR SKFADLTDAA ARNAELLRQA KHEANDYRRQ LQSLTCDLES LRGTNESLER 360
QMREQEERHV REAASYQEAL ARLEEEGQSL KDEMARHLQE YQDLLNVKLA LDIEIATYRK 420
LLEGEENRIT IPVQTFSNLQ IRETSLDTKS VSEGHLKRNI VVKTVEMRDG EVIKESKQEH 480
KDVM 484
Number of amino acids: 484
Molecular weight: 55588.41
Theoretical pI: 5.47
Amino acid composition:
Ala (A) 52 10.7%
Arg (R) 49 10.1%
Asn (N) 17 3.5%
Asp (D) 28 5.8%
Cys (C) 1 0.2%
Gln (Q) 27 5.6%
Glu (E) 63 13.0%
Gly (G) 18 3.7%
His (H) 15 3.1%
Ile (I) 14 2.9%
Leu (L) 58 12.0%
Lys (K) 24 5.0%
Met (M) 16 3.3%
Phe (F) 9 1.9%
Pro (P) 11 2.3%
Ser (S) 28 5.8%
Thr (T) 22 4.5%
Trp (W) 1 0.2%
Tyr (Y) 10 2.1%
Val (V) 21 4.3%
Total number of negatively charged residues (Asp + Glu): 91
Total number of positively charged residues (Arg + Lys): 73
Ext. coefficient 20400
Abs 0.1% (=1 g/l) 0.367, assuming all pairs of Cys residues form cystines
Ext. coefficient 20400
Abs 0.1% (=1 g/l) 0.367, assuming all Cys residues are reduced
1. Bignami A, Eng LF, Dahl D, Uyeda CT. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res. 43:429-35 (1972).
2. Yen SH, Fields KL. Antibodies to neurofilament, glial filament, and fibroblast intermediate filament proteins bind to different cell types of the nervous system. J Cell Biol. 88:115-26 (1981).
3. Shaw G, Osborn M, Weber K. An immunofluorescence microscopical study of the neurofilament triplet proteins, vimentin and glial fibrillary acidic protein within the adult rat brain. Eur. J. Cell Biol. 26:68-82 (1981).
4. Fitch MT, Silver J. CNS injury, glial scars, and inflammation: Inhibitory extracellular matrices and regeneration failure. Exp. Neurol. 209:294-301 (2008).
5. Brenner M, et al. Mutations in GFAP, encoding glial fibrillary acidic protein, are associated with Alexander disease. Nat. Genet. 27:117-20 (2001).
