Overview

  • Product name
  • Description
    Rabbit polyclonal to EAAT1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-FoFr, WB, ICC, Flow Cyt, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
  • Immunogen

    Synthetic peptide, corresponding to 20 residues from the C-terminus of rat EAAT1 (Peptide available as ab127026.)

  • General notes

    Glutamate-aspartate transporter is also known as GLAST.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result, we are pleased to offer this antibody in a purified format as of 23rd June 2017. The following lots are still unpurified and still in stock as of 23rd June 2017 - GR3178513-1,GR3173220-1, GR3173220-2, GR293270-1,  GR320843-2, GR320843-3, GR313192-1, GR320843-1. Lot numbers other than GR3178513-1,GR3173220-1, GR3173220-2, GR293270-1,  GR320843-2, GR320843-3, GR313192-1, GR320843-1 will be purified. Please note that the dilutions may need to be adjusted accordingly. Purified antibodies have the advantage of being enriched for the fraction of immunoglobulin that specifically reacts with the target antigen and for having a reduction of serum proteins.

Properties

Applications

Our Abpromise guarantee covers the use of ab416 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
IHC-FoFr Use at an assay dependent concentration. PubMed: 18714338
WB 1/200 - 1/5000. Predicted molecular weight: 60 kDa.Can be blocked with EAAT1 peptide (ab127026).
ICC Use at an assay dependent concentration.
Flow Cyt 1/10 - 1/200.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    Transports L-glutamate and also L- and D-aspartate. Essential for terminating the postsynaptic action of glutamate by rapidly removing released glutamate from the synaptic cleft. Acts as a symport by cotransporting sodium.
  • Tissue specificity
    Highly expressed in cerebellum, but also found in frontal cortex, hippocampus and basal ganglia.
  • Involvement in disease
    Defects in SLC1A3 are the cause of episodic ataxia type 6 (EA6) [MIM:612656]. EA6 is characterized by episodic ataxia, seizures, migraine and alternating hemiplegia.
  • Sequence similarities
    Belongs to the sodium:dicarboxylate (SDF) symporter (TC 2.A.23) family. SLC1A3 subfamily.
  • Post-translational
    modifications
    Glycosylated.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • EA6 antibody
    • EAA1_HUMAN antibody
    • EAAT1 antibody
    • Excitatory amino acid transporter 1 antibody
    • FLJ25094 antibody
    • GLAST antibody
    • GLAST-1 antibody
    • GLAST1 antibody
    • Glial high affinity glutamate transporter antibody
    • glutamate/aspartate transporter, high affinity, sodium-dependent antibody
    • High affinity neuronal glutamate transporter antibody
    • Slc1a3 antibody
    • Sodium dependent glutamate/aspartate transporter antibody
    • Sodium-dependent glutamate/aspartate transporter 1 antibody
    • Solute carrier family 1 (glial high affinity glutamate transporter) member 3 antibody
    • Solute carrier family 1 member 3 antibody
    see all

Images

  • Anti-EAAT1 antibody (ab416) + Rat brain cortex

    Predicted band size: 60 kDa
    Observed band size: 60 kDa
    Additional bands at: 150 kDa (possible dimer)

  • ab416 staining zebrafish retina sections by IHC-Fr. The tissue was fixed with paraformaldehyde and an antigen retrieval step was performed with sodium citrate pH 6. Blocking of the sample was done with 5%BSA in PBS containing 01% Tween 20 and 0.5% Triton X, for 60 minutes at 23°C, followed by staining with ab416 at 1/100 in blocking solution for 16h at 4°C. An alexa 488 conjugated goat anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei are stained in blue with DAPI. EAAT1 expression can be observed in the inner plexiform layer (in green).

    See Abreview

  • Flow cytometric analysis with ab416 at 1/300 dilution of differentiated human NSCs (green) and undifferentiated stem cells (white). Alexa Fluor© 488 goat anti-rabbit IgG, at 1/500 dilution was used as the secondary antibody.

  • ab416 staining EAAT1 in Chicken retina cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.02% in PBS and blocked with 5% serum for 16 hours at 22°C. Samples were incubated with primary antibody (1/500 in PBS + 0.02% Triton) for 16 hours at 22°C. An undiluted Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab416 at 1/100 dilution staining EAAT1 in mouse coronal tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Adult Sprague–Dawley rats were injected intraperitoneally with pentobarbitone and were transcardially perfused-fixed with heparinized saline and 4% paraformaldehyde in PBS. Brains were removed, were postfixed in 4% paraformaldehyde for 5h, and were stored in PBS. Sections of each brain were pretreated with citrate buffer for 30 min at 65°C to increase antigen retrieval and penetration of the antibodies into the tissues. Sections were permeabilized with 1% Triton X-100 for 5 min and blocked with 3% normal horse serum in 0.1 M PBS, pH 7.4, for 60 min and incubated at room temperature for 48h with primary antibodies. An Alexa Fluor® 488 conjugated anti rabbit was used as secondary at 1/1000 dilution.

  • ab416 (1:500) staining EAAT1 in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of membrane cells in the purkinje glial region .
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ab416 staining EAAT1 in Rat Astrocyte cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 5% Serum for 10 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 1 hour at 25°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal(1/300) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Kia A  et al. Astrocytes expressing ALS-linked mutant FUS induce motor neuron death through release of tumor necrosis factor-alpha. Glia 66:1016-1033 (2018). WB ; Mouse . Read more (PubMed: 29380416) »
  • Johnson J  et al. Valproate and sodium butyrate attenuate manganese-decreased locomotor activity and astrocytic glutamate transporters expression in mice. Neurotoxicology 64:230-239 (2018). Read more (PubMed: 28610743) »
See all 52 Publications for this product

Customer reviews and Q&As

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1-10 of 10 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate ph 6
Permeabilization
No
Specification
brain tissue
Blocking step
Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde

Herr Dr. Markus Kipp

Verified customer

Submitted Sep 07 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain

Abcam user community

Verified customer

Submitted Jul 16 2018

Application
Flow Cytometry
Sample
Human Cell (Differentiated hNSCs)
Permeabilization
Yes - 0.25% Triton X-100 in DPBS
Gating Strategy
Undifferentiated Stem Cells (white)
Specification
Differentiated hNSCs
Preparation
Cell harvesting/tissue preparation method: Accutase
Sample buffer: PBS
Fixation
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 09 2015

Application
Western blot
Sample
Rabbit Tissue lysate - whole (brain)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
brain
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 23 2015

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (Retina, Inner plexiform layer)
Specification
Retina, Inner plexiform layer
Permeabilization
Yes - 0.1% TritonX
Fixative
Paraformaldehyde

Dr. Ryan Macdonald

Verified customer

Submitted Jun 05 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Chicken Cell (retina)
Specification
retina
Fixative
Paraformaldehyde
Permeabilization
Yes - triton X-100 (0.02%) diluted in PBS
Blocking step
Serum as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Mr. Chris Zelinka

Verified customer

Submitted Apr 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Astrocytes)
Specification
Astrocytes
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 19 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (Astrocytes)
Loading amount
30 µg
Specification
Astrocytes
Gel Running Conditions
Reduced Denaturing (12)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (brain)
Specification
brain
Fixative
Formaldehyde
Permeabilization
Yes - 0.02%-Triton
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (primary hippocampal astrocytes)
Loading amount
60 µg
Specification
primary hippocampal astrocytes
Gel Running Conditions
Non-reduced Denaturing (Criterion 4-20% gel)
Blocking step
I-block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Sep 10 2008

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

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