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

  • Methanol fixed human astrocytes (differentiated from H9-derived neuronal) cells labeling EAAT2 using ab41621 at a 1/250 dilution, 1 hr, 22°C (green) followed by an Alexa Fluor® 488 secondary antibody (1/1000 dilution), in ICC/IF.

    Cells were blocked in 5% BSA for 30 mins, 22°C.

  • ICC/IF image of ab41621 stained PC-12 (rat adrenal gland pheochromocytoma cell line) cells.

    The cells were fixed 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:
  • Powell GL  et al. Chronic treatment with N-acetylcysteine decreases extinction responding and reduces cue-induced nicotine-seeking. Physiol Rep 7:e13958 (2019). Read more (PubMed: 30632301) »
  • Cicvaric A  et al. Enhanced synaptic plasticity and spatial memory in female but not male FLRT2-haplodeficient mice. Sci Rep 8:3703 (2018). Read more (PubMed: 29487336) »
See all 28 Publications for this product

Customer reviews and Q&As

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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