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Anti-GFAP antibody [2A5] - Astrocyte Marker (ab4648)

Abreviews (2)Q&A (23)Specific References (7)

Overview

  • Product nameAnti-GFAP antibody [2A5] - Astrocyte MarkerSee all GFAP primary antibodies ...
  • Description
    Mouse monoclonal [2A5] to GFAP - Astrocyte Marker
  • Tested applicationsSandwich ELISA, ICC, IHC-P, ICC/IF, WB, IHC-Fr more 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 controlHomogenized 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 Notes
Sandwich ELISA sELISA: Use a concentration of 0.2 µg/ml. Use this antibody as Capture antibody with Rabbit polyclonal to GFAP - Astrocyte Marker (ab48050) as Detection antibody at 0.5µg/ml.
ICC ICC: Use at an assay dependent dilution.
IHC-P IHC-P: 1/2000.
ICC/IF ICC/IF: 1/10 - 1/50.
WB WB: 1/100 - 1/500. Detects a band of approximately 55 kDa.
IHC-Fr IHC-Fr: Use at an assay dependent dilution.

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.

    • Target information above from: UniProt accession P14136 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
    see all
  • Alternative names
      Astrocyte antibodyFLJ42474 antibodyFLJ45472 antibody
      GFAP antibodyGFAP_HUMAN antibodyGlial Fibrillary Acidic Protein antibodyIntermediate filament protein antibody
    see all

Anti-GFAP antibody [2A5] - Astrocyte Marker images

  • Sandwich ELISA - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Sandwich ELISA - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Standard Curve for GFAP; dilution range 1 pg/ml to 1 ug/ml using Capture Antibody Mouse monoclonal [2A5] to GFAP - Astrocyte Marker (ab4648) at 0.2 ug/ml and Detector Antibody Rabbit polyclonal to GFAP - Astrocyte Marker (ab48050) at 0.5 ug/ml.
  • Immunocytochemistry/ Immunofluorescence - GFAP antibody [2A5] (ab4648)
    Immunocytochemistry/ Immunofluorescence - GFAP antibody [2A5] (ab4648)
    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.
  • Immunocytochemistry/ Immunofluorescence - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Immunocytochemistry/ Immunofluorescence - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Rat neuron/glia cultures stained with mouse monoclonal to GFAP ab4648 (red).
  • - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Rat neuron/glia cultures stained with nmouse monoclonal to GFAP ab 4648 (red).
  • - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Rat neuron/glia cultures stained with mouse monoclonal to GFAP ab4648 (green).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    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.
  • Immunohistochemistry (Frozen sections) - GFAP antibody [2A5] - Astrocyte Marker (ab4648)
    Immunohistochemistry (Frozen sections) - GFAP antibody [2A5] - Astrocyte Marker (ab4648)This image is courtesy of an Abreview submitted by Dr Marian Hruska-Plochan
    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] - Astrocyte Marker (ab4648)

This product has been referenced in:
  • Hao W  et al. Myeloid differentiation factor 88-deficient bone marrow cells improve Alzheimer's disease-related symptoms and pathology. Brain 134:278-92 (2011). ICC/IF ; Mouse . Read more (PubMed: 21115468) »
  • Splinter E  et al. The inactive X chromosome adopts a unique three-dimensional conformation that is dependent on Xist RNA. Genes Dev 25:1371-83 (2011). Read more (PubMed: 21690198) »

See all 7 Publications for this product

Publishing research using ab4648 ? Please let us know so that we can cite the reference in this datasheet

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Displaying 1 - 10 of 25 results for Abreviews and Q&A

10
Votes: Highest First

IHC-Fr with rat retina works very well
same protocol with mouse retina does not work: no staining
Ab: 1/200
block: NGS
2nd: same as for rat, works well
tried several untreated and treated sections

A free of charge replacement was sent.

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFAP antibody [2A5] - Astrocyte Marker

 Excellent
Abreviews
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
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Username

Verified customer

Submitted Dec 26 2012

Inquiry: Hello Sir/Madam, I am a graduate student looking for a product that can differentiate between astrocytes and neurons in a dissociated neuronal cell culture from mouse brain. I have never done GFAP staining before and I cannot find a straight forw...

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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 it...

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Application: sELISA
Capture ab: ab4648, ELISA coating buffer, ph 9.9, polystyrene plate, 0.2 ug/mL O/N @ 4oC
Wash twice with PBS
Block (PBS + 5% goat serum) 90 min. @ RT. Tried 3% BSA as well
Introduce antigen: 2 hr. RT up to 100 ng/mL (wi...

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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 bi...

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Dear Concerning,

The user will use "Anti-GFAP antibody [2A5] - Astrocyte Marker (ab4648)" also on rat cell culture by IF. The antibody include "10mM Sodium Azide" as a preservative. So, is it suitable to use on live cell sta...

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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 resis...

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Product: Anti-MAP2 antibody (ab70218)
Test: on rat cell culture by ICC / IF

P.S: will use "Anti-GFAP antibody [2A5] - Astrocyte Marker (ab4648)" also on rat cell culture by IF. The antibody include "10mM Sodium Azide" a...

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Thank you for contacting us,

For ab70218:
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I'm wanting to immunostain protoplasmic astrocytes in human brain paraffin sections.

Can you tell me which of your antibodies would be best for this? Is there any published work in this area.

Thanks,

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.0038243 ...

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We attempted to stain frozen spinal cord tissue specimens fixed in either paraformaldehyde or ethanol using the anti-GFAP antibody ab4648 without any luck. After reviewing previous concerned posts regarding this antibody, I have to confess I'm a little co...

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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 product ...

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Can you please tell me the protocol used to validate ab4648 and ab48050 as a sandwich ELISA pair?

Thank you for your patience. The protocol used was as follows:
Buffers and Reagents:
Carbonate Coating Buffer (100 mM)
Antigen or antibody should be diluted in coating buffer to immobilize them to the wells:
3.03 g Na2CO3,
6.0 g N...

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I am using this antibody along with ab48050 in a sELISA experiment. The detection antibody ab48050 seems to work well alone, but I cannot detect a signal combined with ab4648. Can I try another aliquot?

Thank you for contacting Abcam regarding ab4648.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status ...

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