Overview

  • Product nameAnti-GFAP antibody [2A5]See all GFAP primary antibodies ...
  • Description
    Mouse monoclonal [2A5] to GFAP
  • Tested applicationsSandwich ELISA, ICC, IHC-P, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Pig
    Predicted to work with: all Mammals

    Does not react with

    Chicken
  • Immunogen

    A preparation of purified pig spinal core GFAP.

  • Positive control
    • Homogenized cow, pig, human, rat, mouse or other mammalian brain extract in 1%SDS, 6M urea.

Properties

Applications

Our Abpromise guarantee covers the use of ab4648 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA 1/5000.

Use this antibody as Capture antibody with Rabbit polyclonal to GFAP - Astrocyte Marker (ab48050) as Detection antibody at 0.5µg/ml.

ICC Use at an assay dependent concentration.
IHC-P 1/2000.
ICC/IF 1/10 - 1/50.
WB 1/100 - 1/500. Detects a band of approximately 55 kDa.
IHC-Fr Use at an assay dependent concentration.

Target

  • FunctionGFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells.
  • Tissue specificityExpressed in cells lacking fibronectin.
  • Involvement in diseaseDefects in GFAP are a cause of Alexander disease (ALEXD) [MIM:203450]. Alexander disease is a rare disorder of the central nervous system. It is a progressive leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers which are cytoplasmic inclusions in astrocytes. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death usually within the first decade. Infants with Alexander disease develop a leukoencephalopathy with macrocephaly, seizures, and psychomotor retardation. Patients with juvenile or adult forms typically experience ataxia, bulbar signs and spasticity, and a more slowly progressive course.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • Post-translational
    modifications
    Phosphorylated by PKN1.
  • Cellular localizationCytoplasm. Associated with intermediate filaments.
  • Information by UniProt
  • Database links
  • Alternative names
    • Astrocyte antibody
    • FLJ42474 antibody
    • FLJ45472 antibody
    • GFAP antibody
    • GFAP_HUMAN antibody
    • Glial Fibrillary Acidic Protein antibody
    • Intermediate filament protein antibody
    see all

Anti-GFAP antibody [2A5] images

  • Standard Curve for GFAP; dilution range 1 pg/ml to 1 µg/ml using Capture Antibody Mouse monoclonal [2A5] to GFAP - Astrocyte Marker (ab4648) at 1/5000 and Detector Antibody Rabbit polyclonal to GFAP - Astrocyte Marker (ab48050) at 0.5 µg/ml.

  • Rat cortical neurons and glia in mixed tissue culture stained with Chicken polyclonal to MAP2 - ab5392 (green) at 1/30000 and Mouse monoclonal to GFAP - ab4648 (red) at 1/100. Nuclei of all cells are stained with Hoechst dye (blue). Picture taken with a Zeiss 20X objective and documented with a Digital SPOT camera.
  • Rat neuron/glia cultures stained with mouse monoclonal to GFAP ab4648 (red).
  • Rat neuron/glia cultures stained with nmouse monoclonal to GFAP ab 4648 (red).
  • Rat neuron/glia cultures stained with mouse monoclonal to GFAP ab4648 (green).
  • Ab4648 staining human substantia nigra. Staining is localised to the cytoplasm.
    Left panel: with primary antibody diluted 1:4000. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ab4648 staining GFAP - Astrocyte Marker in pig brain tissue sections by IHC-Fr (formaldehyde-fixed Frozen sections). Tissue was fixed with formaldehyde (4%), permeabilized with Triton X-100 and blocked with 0.2% milk for 30 minutes at 24°C. Samples were incubated with primary antibody 1/2000 (TBS + 1% Triton X-100 + 0.2% milk) for 24 hours at 4°C. A Biotin-conjugated Sheep polyclonal to mouse IgG, dilution 1/400, was used as secondary antibody. Endogenous peroxidase was blocked with 0.3% H2O2 in methanol for 20 minutes.

    See Abreview

References for Anti-GFAP antibody [2A5] (ab4648)

This product has been referenced in:

See all 9 Publications for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Brain)
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 08 2013


The antibodies mentioned above are suitable for Rat as well as human samples. ab9018 is yet to be tested in IHC-Fr however as it is tested in IHC-FoFr so it is more likely to work.

The suitable secondary's are for ab4648, ab82259 and ab90...

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A free of charge replacement was sent.

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Embryonic rat cerebral cortex)
Specification Embryonic rat cerebral cortex
Fixative Paraformaldehyde
Permeabilization Yes - Ice cold methanol
Blocking step Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 26 2012

Thank you for contacting Abcam.

When comparing multiple products that have been tested in the same species and applications, we like to use the abreviews and specific references to judge the potential for an antibody to perform as indicated on...

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Thank you for contacting Abcam.

Earlier we discussed your sELISA with ab4648 as the capture and ab48050 as the detector. As we discussed, since you would like to use your streptavidin-gold conjugated secondary, a detector that is conjugated to...

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Thank you for contacting us.

There is a small amount of azide in the preparation, but I don't think that would be a problem for two reasons.

1. the antibody can be greatly diluted
2. cells in tissue culture are surprisingly re...

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Thank you for contacting us,

For ab70218:
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For ab22623:
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For ab81887:
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Thank you for contacting us.

After completing a short literature search, GFAP is commonly used as an protoplasmic astrocyte marker. Below are two references:

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.00382...

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Thank you for taking time tocontact us. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring produ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"