Overview

  • Product name
  • Description
    Rabbit polyclonal to EAAT2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat
    Predicted to work with: Human
  • Immunogen

    Synthetic peptide within Rat EAAT2 aa 550 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    Database link: P31596
    (Peptide available as ab41752)

  • Positive control
    • WB: Mouse and rat brain tissue lysates. IHC-Fr: Mouse brain tissue. ICC/IF: PC-12 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab41621 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).
ICC/IF Use a concentration of 5 µg/ml.
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.
  • Sequence similarities
    Belongs to the sodium:dicarboxylate (SDF) symporter (TC 2.A.23) family. SLC1A2 subfamily.
  • Post-translational
    modifications
    Glycosylated.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • EAA2_HUMAN antibody
    • EAAT2 antibody
    • Excitatory amino acid transporter 2 antibody
    • Excitotoxic amino acid transporter 2 antibody
    • Glial high affinity glutamate transporter antibody
    • GLT 1 antibody
    • GLT1 antibody
    • Glutamate aspartate transporter II antibody
    • Glutamate transporter 1 antibody
    • Glutamate/aspartate transporter II antibody
    • Slc1a2 antibody
    • Sodium dependent glutamate aspartate transporter 2 antibody
    • Sodium-dependent glutamate/aspartate transporter 2 antibody
    • solute carrier family 1 (glial high affinity glutamate transporter), member 2 antibody
    • Solute carrier family 1 glial high affinity glutamate transporter member 2 antibody
    • Solute carrier family 1 member 2 antibody
    see all

Images

  • All lanes : Anti-EAAT2 antibody (ab41621) at 1 µg/ml

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 62 kDa
    Observed band size: 62 kDa

  • Immunohistochemical staining of PFA-fixed, frozen mouse brain tissue using undiluted ab41621. Tissue sections were permeabilized using Triton-X100 and incubated with ab41621 for 12 hours at 4°C. ab150077 (goat anti-rabbit IgG H&L Alexa Fluor® 488) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab41621 stained PC12 (rat adrenal gland pheochromocytoma cell line) cells. The cells were fied in 4% formaldehyde (10 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab41621, 5 µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

References

This product has been referenced in:
  • 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) »
  • Zoltowska KM  et al. Novel interaction between Alzheimer's disease-related protein presenilin 1 and glutamate transporter 1. Sci Rep 8:8718 (2018). IP ; Mouse . Read more (PubMed: 29880815) »
See all 26 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (rat brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: tri-sodium citrate buffer
Permeabilization
Yes - triton 0.3%
Specification
rat brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10 · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Bartosz Pomierny

Verified customer

Submitted Mar 15 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Cow Tissue lysate - whole (Muscle)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
Muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 25 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Spinal cord)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
20 µg
Specification
Spinal cord
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 29 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain tissue)
Permeabilization
Yes - Triton-X100
Specification
Brain tissue
Fixative
Paraformaldehyde

Dr. Daniel Berg

Verified customer

Submitted Aug 20 2015

Question
Answer

Thank you for contacting us.
I have contacted the Abreview customer regarding more information about the staining with ab41621. I will let you know if I hear back from her.
As for ab61859, I have the following IHC-P protocol:
General IHC Protocol
Paraffin Sections:
1. Deparaffinize sections in xylene, 2x5min.
2. Hydrate with 100% ethanol, 2x3min.
3. Hydrate with 95% ethanol, 1min.
4. Rinse in distilled water.
Tissue Staining:
1. Follow procedure for pretreatment as required.
2. Procedure for Immunoenzyme Staining.
3. Rinse Sections in Washing Buffer (PBS+0.1% Triton X-100, pH 7.4) for 2x2 min.
4. Serum Blocking: incubate sections in normal serum block – species same as secondary antibody( Wash Buffer + 5% Normal Sera). Note: since this protocol uses avidin-biotin detection system, avidin/biotin block may be needed based on tissue type. If you do, the avidin/biotin blocking should be done after normal serum block and before primary antibody incubation.
5. Primary Antibody: incubate sections in primary antibody at appropriate dilution in Wash Buffer for 1 hour at room temperature or overnight at 4C. Note:Do not rinse sections between serum block and primary antibody incubation.
6. Rinse in washing buffer for 3x2 min.
7. Peroxidase Blocking: incubate sections in peroxidase blocking solution for 10 minutes at room temperature.
8. Rinse in washing buffer for 3x2 min.
9. Secondary Antibody: incubate sections in biotinylated secondary antibody at an appropriate dilution in Wash Buffer for 30 minutes at room temperature.
10. Rinse in washing buffer for 3x2 min.
11. Detection: incubate sections in streptavidin-HRP in Wash Buffer for 30 minutes at room temperature.
12. Rinse in washing buffer for 3x2 min.
13. Chromagen/Substrate: incubate sections in peroxidase substrate solution.
14. Rinse in washing buffer for 3x2 min.
15. Rinse in running tap water for 2-5 minutes.
16. Dehydrate through 95% ethanol for 1 minute, 100% ethanol for 2x3min.
17. Clear in xylene for 2x5min.
18. Coverslip with mounting medium.

Peroxidase Block:
0.3% H2O2 in PBS (for Paraffin Sections)
0.3% H2O2 in Methanol (for Frozen Sections)

Avidin/Biotin Block
0.001% Avidin in PBS
0.001% Biotin in PBS
Block for 10 minutes with first solution, rinse with PBS, block for 10 minutes with second solution.
I am checking with the lab regarding the antigen retrieval method used with this antibody.
I hope this information is so far helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain sections)
Specification
Brain sections
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No

Dr. Sophie Pezet

Verified customer

Submitted Jan 25 2010

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

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