Anti-GFAP antibody ValidAbTM

(HB6406)
Technical documents: SDS Datasheet

Product overview

Name Anti-GFAP antibody ValidAbTM
Host Chicken
Clonality Polyclonal
Target GFAP
Description

Antibody to GFAP - cytoskeletal protein used as an astrocyte marker. Part of the ValidAb™ range of highly validated, data-rich antibodies.

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Validation data

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

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.

Open data link

Please follow this link to OSF

Target information

UniProt ID P14136
Structure image  Chemical Structure
Gene name GFAP
NCBI full gene name glial fibrillary acidic protein
Entrez gene ID

2670

Amino acids

432 (49.9kDa)

Isoforms

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

FAQs

References for Anti-GFAP antibody ValidAbTM

References are publications that support the biological activity of the product
  • Importance of GFAP isoform-specific analyses in astrocytoma.

    van Bodegraven EJ et al (2019) Glia 67 : 1417-1433
  • The role of GFAP and vimentin in learning and memory.

    Wilhelmsson U et al (2019) Biological chemistry 400 : 1147-1156
  • GFAP-expressing progenitors are the principal source of constitutive neurogenesis in adult mouse forebrain.

    Garcia AD et al (2004) Nature neuroscience 7 : 1233-41
  • Glial fibrillary acidic protein: GFAP-thirty-one years (1969-2000).

    Eng LF et al (2000) Neurochemical research 25 : 1439-51
  • GFAP and astrogliosis.

    Eng LF et al (1994) Brain pathology (Zurich, Switzerland) 4 : 229-37

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