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    eaat3-antibody-epr6774b-ab124802.pdf

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Neuroscience Neurotransmitter Amino Acids Glutamate
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RecombinantRabMAb

Recombinant Anti-EAAT3 antibody [EPR6774(B)] (ab124802)

  • Datasheet
Reviews (3) Submit a question References (9)

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Western blot - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
  • Western blot - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
  • OI-RD Scanning - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
  • Anti-EAAT3 antibody [EPR6774(B)] (ab124802)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6774(B)] to EAAT3
  • Suitable for: IHC-P, WB
  • Reacts with: Human

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Overview

  • Product name

    Anti-EAAT3 antibody [EPR6774(B)]
    See all EAAT3 primary antibodies
  • Description

    Rabbit monoclonal [EPR6774(B)] to EAAT3
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human EAAT3 aa 150-250. The exact sequence is proprietary.

  • Positive control

    • WB: Human fetal brain, human brain and human testis lysates . IHC-P: Human kidney tissue, and human glioma tissue. Flow Cyt: U-87 MG cells.
  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 3.80 x 10 -12 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6774(B)
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmitter
    • Amino Acids
    • Glutamate
    • Neuroscience
    • Neurotransmitter
    • Amino Acids
    • Aspartate
    • Neuroscience
    • Neurotransmitter
    • Transporters
    • Glutamate

Associated products

  • Alternative Versions

    • HRP Anti-EAAT3 antibody [EPR6774(B)] (ab203479)
    • Anti-EAAT3 antibody [EPR6774(B)] - BSA and Azide free (ab240000)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124802 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
WB
Human
Application Abreviews Notes
IHC-P (1)
1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (1)
1/10000 - 1/50000.

Detects a band of approximately 70 kDa (predicted molecular weight: 57 kDa).

Notes
IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/10000 - 1/50000.

Detects a band of approximately 70 kDa (predicted molecular weight: 57 kDa).

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • 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. Negatively regulated by ARL6IP5.
    • Tissue specificity

      Expressed in all tissues tested including liver, muscle, testis, ovary, retinoblastoma cell line, neurons and brain (in which there was dense expression in substantia nigra, red nucleus, hippocampus and in cerebral cortical layers).
    • Sequence similarities

      Belongs to the sodium:dicarboxylate (SDF) symporter (TC 2.A.23) family. SLC1A1 subfamily.
    • Post-translational
      modifications

      Glycosylated.
    • Cellular localization

      Membrane.
    • Target information above from: UniProt accession P43005 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 6505 Human
      • Omim: 133550 Human
      • SwissProt: P43005 Human
      • Unigene: 444915 Human
      • Alternative names

        • EAA3_HUMAN antibody
        • EAAC 1 antibody
        • EAAC1 antibody
        • EAAT 3 antibody
        • Excitatory amino acid carrier 1 antibody
        • Excitatory amino acid carrier1 antibody
        • Excitatory amino acid transporter 3 antibody
        • Excitatory amino acid transporter3 antibody
        • Excitatory amino-acid carrier 1 antibody
        • GLUTAMATE TRANSPORTER, HIGH-AFFINITY antibody
        • MEAAC 1 antibody
        • MEAAC1 antibody
        • Neuronal and epithelial glutamate transporter antibody
        • REAAC 1 antibody
        • REAAC1 antibody
        • Slc1 a1 antibody
        • Slc1a 1 antibody
        • SLC1A1 antibody
        • Sodium dependent glutamate/aspartate transporter 3 antibody
        • Sodium-dependent glutamate/aspartate transporter 3 antibody
        • Solute carrier family 1 (neuronal / epithelial high affinity glutamate transporter, system Xag), member 1 antibody
        • SOLUTE CARRIER FAMILY 1 (NEURONAL/EPITHELIAL HIGH AFFINITY GLUTAMATE TRANSPORTER), MEMBER 1 antibody
        • Solute carrier family 1 (neuronal/epithelial high affinity glutamate transporter, system Xag), member 1 antibody
        • Solute carrier family 1 member 1 antibody
        • Solute carrier family 1, member 1 antibody
        see all

      Images

      • Western blot - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Western blot - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        All lanes : Anti-EAAT3 antibody [EPR6774(B)] (ab124802) at 1/50000 dilution ((unpurified))

        Lane 1 : Human fetal brain tissue lysate
        Lane 2 : Human testis tissue lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

        Observed band size: 70 kDa
        why is the actual band size different from the predicted?


        Exposure time: 30 seconds


        Blocking and Diluting buffer : 5% NFDM/TBST

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labeling EAAT3 with purified ab124802 at 1/1000 dilution (0.8 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

      • Western blot - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Western blot - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Anti-EAAT3 antibody [EPR6774(B)] (ab124802) at 1/1000 dilution (Purified) + Human brain lysates at 15 µg

        Secondary
        Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

        Observed band size: 70 kDa why is the actual band size different from the predicted?



        We are unsure how to define the extra bands.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)

        Unpurified ab124802, at 1/500 dilution, staining EAAT3 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • OI-RD Scanning - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        OI-RD Scanning - Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD
      • Anti-EAAT3 antibody [EPR6774(B)] (ab124802)
        Anti-EAAT3 antibody [EPR6774(B)] (ab124802)

      Protocols

      • Immunohistochemistry protocols
      • Immunocytochemistry & immunofluorescence protocols
      • Western blot protocols

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
    • References (9)

      Publishing research using ab124802? Please let us know so that we can cite the reference in this datasheet.

      ab124802 has been referenced in 9 publications.

      • Liu XX  et al. Endothelial Cdk5 deficit leads to the development of spontaneous epilepsy through CXCL1/CXCR2-mediated reactive astrogliosis. J Exp Med 217:N/A (2020). PubMed: 31699822
      • Hövel FFV  et al. FGF-2 isoforms influence the development of dopaminergic neurons in the murine substantia nigra, but not anxiety-like behavior, stress susceptibility, or locomotor behavior. Behav Brain Res 374:112113 (2019). PubMed: 31381976
      • Biolley C  et al. Punicalagin increases glutamate absorption in differentiated Caco-2 cells by a mechanism involving gene expression regulation of an EAAT3 transporter. Food Funct 10:5333-5338 (2019). PubMed: 31389458
      • Garza-Lombó C  et al. Systemic L-buthionine-S-R-sulfoximine administration modulates glutathione homeostasis via NGF/TrkA and mTOR signaling in the cerebellum. Neurochem Int 121:8-18 (2018). PubMed: 30300680
      • González LF  et al. Neurochemical and behavioral characterization of neuronal glutamate transporter EAAT3 heterozygous mice. Biol Res 50:29 (2017). PubMed: 28927446
      • Huang C  et al. Estrogen regulates excitatory amino acid carrier 1 (EAAC1) expression through sphingosine kinase 1 (SphK1) transacting FGFR-mediated ERK signaling in rat C6 astroglial cells. Neuroscience 319:9-22 (2016). PubMed: 26804240
      • Beaudin S & Welsh J 1,25-Dihydroxyvitamin D induces the glutamate transporter SLC1A1 and alters glutamate handling in non-transformed mammary cells. Mol Cell Endocrinol 424:34-41 (2016). WB ; Human . PubMed: 26774511
      • Ramos-Chávez LA  et al. Neurological effects of inorganic arsenic exposure: altered cysteine/glutamate transport, NMDA expression and spatial memory impairment. Front Cell Neurosci 9:21 (2015). WB ; Mouse . PubMed: 25709567
      • Martinez-Lozada Z  et al. Activation of sodium-dependent glutamate transporters regulates the morphological aspects of oligodendrocyte maturation via signaling through calcium/calmodulin-dependent kinase IIß's actin-binding/-stabilizing domain. Glia 62:1543-58 (2014). PubMed: 24866099

      Customer reviews and Q&As

      Show All Reviews Q&A
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      1-3 of 3 Abreviews or Q&A

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EAAT3 antibody [EPR6774(B)]

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Dog Tissue sections (cruciate ligament, meniscus and brain)
      Antigen retrieval step
      Enzymatic - Buffer/Enzyme Used: trypsin from porcine pancreas
      Permeabilization
      No
      Specification
      cruciate ligament, meniscus and brain
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
      Fixative
      Formaldehyde
      Read More

      Dr. Joel Alves

      Verified customer

      Submitted Feb 11 2019

      Western blot abreview for Anti-EAAT3 antibody [EPR6774(B)]

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (Human umbilical vein endothelial cell)
      Gel Running Conditions
      Non-reduced Denaturing (10% gel)
      Loading amount
      20 µg
      Specification
      Human umbilical vein endothelial cell
      Blocking step
      Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      Submitted Aug 01 2017

      Immunocytochemistry/ Immunofluorescence abreview for Anti-EAAT3 antibody [EPR6774(B)]

      Excellent
      Abreviews
      Abreviews
      This product is known to not work in this application or species.
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Mouse Cell (Brain)
      Permeabilization
      No
      Specification
      Brain
      Blocking step
      Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
      Fixative
      Paraformaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted May 12 2017

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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