Figure 11. Astrocytes stained for GFAP with HB6406 in the cerebellum
Rat cerebellum stained by HB6406 for GFAP. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before ...View more
1/11
Figure 1. An astrocyte surrounded by neurons in a cultured rat neuron preparation.
HB6406 stains GFAP in the star shaped projections of an astrocyte surrounded by neurofilament L stained with HB7266 (rabbit monoclonal). Method: neurons were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 3 ...View more
A section of hippocampal CA1 stained by HB6406 for GFAP (chicken polyclonal), HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections ...View more
Rat hippocampus stained by HB6406 for GFAP (chicken polyclonal), HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubat ...View more
1/11
Figure 4. Populations of astrocytes and neurons in a cultured rat neuron preparation.
HB6406 stains the GFAP positive glia that surround and support the neurofilament L expressing neurones (HB7266 stained: rabbit monoclonal) in a rat cultured neuron preparation. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Trito ...View more
1/11
Figure 5. Concentration response of HB6406 staining in rat cortex and cerebellum.
HB6406 produces strong staining of astrocytes in cortex and cerebellum at dilutions as low at 1:8,000. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were ...View more
1/11
Figure 6. Concentration response of HB6406 staining in cultured rat neurons.
HB6406 produces strong staining of astrocytes in a cultured neuron preparation at dilutions as low at 1:8,000. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 300mM glycine. HB6406 (chi ...View more
1/11
Figure 7. GFAP expression in various tissue lysates and preparations.
HB6406 revealed the ≈55kDa band associated with GFAP only in neural tissue samples. Method: mouse brain and rat brain membrane (P2) and cytosol fractions were prepared following previous work (Molnar et al., 1993. Neuroscience 53:307-326) from freshly collected adult brains. Other tissue lysates were prepared following established protocols from freshly dissected tissue (see ou ...View more
1/11
Figure 8. Concentration response of HB6406 staining in a rat brain cytosol preparation.
HB6406 shows strong affinity for GFAP with bands visible at as low as a 1 in 128,000 dilution. Method: cytosol fractions were prepared from fresh rat brains following established protocols (Molnar et al., 1993. Neuroscience 53:307-326). Rat cytosol samples were loaded (20µg / lane) onto a 12% acrylamide gel alongside a protein ladder (NEB Prestained protein standard, P7718S) be ...View more
1/11
Figure 9. Independent antibody validation of HB6406 and HB8001 in a cultured rat neuron preparation.
HB6406 and HB8001 staining completely overlaps in a cultured neuron preparation therefore showing strong evidence for specificity. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 300mM ...View more
1/11
Figure 10. Populations of astrocytes and neurones in a cultured rat neuron preparation.
HB6406 stains the GFAP positive glia that surround and support the neurofilament L expressing neurones (HB7266 stained: rabbit monoclonal) in a rat cultured neuron preparation. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Trito ...View more
1/11
Figure 11. Astrocytes stained for GFAP with HB6406 in the cerebellum
Rat cerebellum stained by HB6406 for GFAP. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before ...View more
Figure 1. An astrocyte surrounded by neurons in a cultured rat neuron preparation.
HB6406 stains GFAP in the star shaped projections of an astrocyte surrounded by neurofilament L stained with HB7266 (rabbit monoclonal). Method: neurons were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 3 ...View more
A section of hippocampal CA1 stained by HB6406 for GFAP (chicken polyclonal), HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections ...View more
Rat hippocampus stained by HB6406 for GFAP (chicken polyclonal), HB6457 for Parvalbumin (mouse monoclonal) and HB6498 for NeuN (rabbit polyclonal). Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubat ...View more
Figure 4. Populations of astrocytes and neurons in a cultured rat neuron preparation.
HB6406 stains the GFAP positive glia that surround and support the neurofilament L expressing neurones (HB7266 stained: rabbit monoclonal) in a rat cultured neuron preparation. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Trito ...View more
Figure 5. Concentration response of HB6406 staining in rat cortex and cerebellum.
HB6406 produces strong staining of astrocytes in cortex and cerebellum at dilutions as low at 1:8,000. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were ...View more
Figure 6. Concentration response of HB6406 staining in cultured rat neurons.
HB6406 produces strong staining of astrocytes in a cultured neuron preparation at dilutions as low at 1:8,000. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 300mM glycine. HB6406 (chi ...View more
Figure 7. GFAP expression in various tissue lysates and preparations.
HB6406 revealed the ≈55kDa band associated with GFAP only in neural tissue samples. Method: mouse brain and rat brain membrane (P2) and cytosol fractions were prepared following previous work (Molnar et al., 1993. Neuroscience 53:307-326) from freshly collected adult brains. Other tissue lysates were prepared following established protocols from freshly dissected tissue (see ou ...View more
Figure 8. Concentration response of HB6406 staining in a rat brain cytosol preparation.
HB6406 shows strong affinity for GFAP with bands visible at as low as a 1 in 128,000 dilution. Method: cytosol fractions were prepared from fresh rat brains following established protocols (Molnar et al., 1993. Neuroscience 53:307-326). Rat cytosol samples were loaded (20µg / lane) onto a 12% acrylamide gel alongside a protein ladder (NEB Prestained protein standard, P7718S) be ...View more
Figure 9. Independent antibody validation of HB6406 and HB8001 in a cultured rat neuron preparation.
HB6406 and HB8001 staining completely overlaps in a cultured neuron preparation therefore showing strong evidence for specificity. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Triton X-100 followed by blocking in 1% BSA, 300mM ...View more
Figure 10. Populations of astrocytes and neurones in a cultured rat neuron preparation.
HB6406 stains the GFAP positive glia that surround and support the neurofilament L expressing neurones (HB7266 stained: rabbit monoclonal) in a rat cultured neuron preparation. Method: neurones were cultured from E17-E18 rat embryos following established protocols (Martin and Henley, 2004. EMBO, 4749–4759) and fixed with 4% PFA on DIV21. Cells were permeabilised with 0.1% Trito ...View more
Figure 11. Astrocytes stained for GFAP with HB6406 in the cerebellum
Rat cerebellum stained by HB6406 for GFAP. Method: Rat brains were dissected and fixed overnight in 4% PFA before then being incubated in 30% sucrose (in PBS) until sunk (approx. 48hrs). A freezing microtome was used to cut 40µm horizontal slices before sections were incubated in 1% NaBH4 for 30 minutes. Sections were blocked in 0.05M glycine, 2% BSA and 3% donkey serum before ...View more
Product information
Immunogen
Recombinant human GFAP (isoform 1) expressed in and purified from E. coli
Isotype
IgY
Purification
Unpurified
Formulation
Lyophilised. When reconstituted contains IgY preparation with 5mM sodium azide and 1% recombinant BSA.
Predicted species reactivity
Mouse, Rat, Human, Pig, Horse, Cow
Tested species reactivity
Mouse, Rat
Tested applications
Applications
ICC, WB, IHC(IF)
Western blot optimal concentration
1:8,000 dilution as tested in a rat brain cytosol preparation.
IHC(IF) optimal concentration
1:4,000 dilution as tested in free-floating paraformaldehyde fixed horiztonal rat brain sections.
ICC optimal concentration
1:8,000 dilution as tested in cultured rat neurones.
Positive control
GFAP is highly expressed in neural tissues containing astrocytes. It is not widely expressed in cell lines, however it is in specific lines such as U-87 MG.
Negative control
Most non-neural tissues. Please note that GFAP expression has been reported in a subset of pancreatic and hepatic cells in rats and mice kidney cells. It is generally poorly expressed in common cell lines such as HeLa or HEK293.
GFAP has three confirmed and 21 potential isoforms. Isoform 1 (GFAP alpha): canonical, 49.9kDa; Isoform 2 (GFAP epsilon): amino acid changes between positions 391 and 432, 49.5kDa; Isoform 3 (GFAP kappa): amino acid changes between positions 391 and 432, 50.3kDa
Expression
GFAP is primarily expressed within astrocytes of the central nervous system alongside also expressing in non-myelinating Schwann cells of the peripheral nervous system and satellite cells of the peripheral ganglia. GFAP expression has also been reported in Leydig cells of the testis alongside stellate cells from the pancreas and liver in rats.
Subcellular expression
GFAP is a key cytoskeletal component therefore is widely expressed as bundles of GFAP positive fibres.
Processing
Following translation, no processing is required for GFAP to reach its active conformation.
Post translational modifications
GFAP is subjected to numerous post-translational modifications including 9 phosphorylation sites which are the target of AURKB and ROCK1 alongside 5 separate citrullination sites.
Homology (compared to human)
Rat, mouse and human GFAP proteins have a 90% similarity score in a direct BLAST comparison.
Similar proteins
Other type III intermediate filament proteins have homology with GFAP including Vimentin (58%), Desmin (59%) and Peripherin (56%) when assessed using BLAST.
Storage & Handling
Storage instructions
-20°C then use reconstitution advice
Handling
Upon receipt store at either -20°C or -80°C. When ready to use there are three options:
Reconstitute with 100μl dH2O and store at 4°C
Reconstitue with 50μl dH2O and 50μl glycerol then store at -20°C
Reconstitue with 100μl dH2O, aliquot then snap freeze and store at -80°C
For more information read our guide on the best care for your product. Take care when opening as the precipitate is extremely light and can easily be lost if disturbed. When reconstituting make sure that the antibody is thoroughly dissolved by pipetting up and down before giving the antibody a brief spin at 10,000g to make sure that all material is recovered and at the bottom of the tube.
Important
This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use
What guarantee do you have that my GFAP antibody will perform as expected?
We guarantee that your GFAP antibody will work for the applications and species we list on the datasheet. If the antibody fails to perform as expected then we are happy to offer a 100% refund guarantee. For more details please see our guarantee policy.
Will my GFAP antibody work against species that have not been listed on the datasheet?
A species not being listed doesn’t mean that the GFAP antibody won’t work, just that we haven’t tested it. If you test one of our antibodies in a new species please let us know (positive or negative)!
What protocols are available for use with this GFAP antibody
We have made a comprehensive collection of protocols that we have used in our experiments to validate this GFAP antibody.
Antibody to Calbindin - calcium binding protein used as a marker for an inhibitory interneuron subtype. Part of the ValidAb™ range of highly validated, data-rich antibodies.