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EAAT1 Recombinant Rabbit Monoclonal Antibody [JA30-35] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750415

EAAT1 Recombinant Rabbit Monoclonal Antibody [JA30-35] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IHC-Fr

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Predicted reactivity

  • Cynomolgus monkey

  • Pig

Predicted species support after-sales service
Conjugation

  • unconjugated

Overview

Product Name

EAAT1 Recombinant Rabbit Monoclonal Antibody [JA30-35] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human EAAT1 aa 171-220 / 542.

Species Reactivity

Human, Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

Predicted species support after-sales service

Validated Applications

WB, IHC-P, IF-Tissue, IHC-Fr

Molecular Weight

Predicted band size: 60 kDa

Positive Control

Human cerebellum tissue, mouse cerebellum tissue, rat cerebellum tissue, Mouse brain tissue lysate, Mouse cerebellum tissue lysate, Rat brain tissue lysate, Rat cerebellum tissue lysate.

Conjugation

unconjugated

Clone Number

JA30-35

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:1,000-1:8,000

  • IF-Tissue

  • 1:500

  • IHC-Fr

  • 1:500

Target

Function

Sodium-dependent, high-affinity amino acid transporter that mediates the uptake of L-glutamate and also L-aspartate and D-aspartate. Functions as a symporter that transports one amino acid molecule together with two or three Na+ ions and one proton, in parallel with the counter-transport of one K+ ion. Mediates Cl- flux that is not coupled to amino acid transport; this avoids the accumulation of negative charges due to aspartate and Na+ symport. Plays a redundant role in the rapid removal of released glutamate from the synaptic cleft, which is essential for terminating the postsynaptic action of glutamate. This gene encodes a member of a member of a high affinity glutamate transporter family. This gene functions in the termination of excitatory neurotransmission in central nervous system. Mutations are associated with episodic ataxia, Type 6. Alternative splicing results in multiple transcript variants.

Background References

1. Xu NJ et al. Morphine withdrawal increases glutamate uptake and surface expression of glutamate transporter GLT1 at hippocampal synapses. J Neurosci. 23(11): 4775-4784 (2003).

2. Ueda, Hideho et al. Caveolin-1 Localization in Müller Cells of the Retina ACTA HISTOCHEMICA ET CYTOCHEMICA. 35: 423-428 (2002).

Sequence Similarity

Belongs to the dicarboxylate/amino acid:cation symporter (DAACS) (TC 2.A.23) family. SLC1A3 subfamily.

Tissue Specificity

Detected in brain. Detected at very much lower levels in heart, lung, placenta and skeletal muscle. Highly expressed in cerebellum, but also found in frontal cortex, hippocampus and basal ganglia.

Post-translational Modification

Glycosylated.

Subcellular Location

Cell membrane.

UNIPROT

SwissProt: P43003 Human

SwissProt: P56564 Mouse

SwissProt: P24942 Rat

Synonyms

GLAST-1

SLC1A3

EAAT1

GLAST

GLAST1

Images

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: Cerebellum<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1:500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: Cerebellum

    Sample: Frozen section

    Antibody concentration: 1:500

    Antigen retrieval: Not required

  • Application: IHC-Fr<br /><br />Species: Rat<br /><br />Site: Cerebellum<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1:500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Rat

    Site: Cerebellum

    Sample: Frozen section

    Antibody concentration: 1:500

    Antigen retrieval: Not required

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: Cerebellum<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1:500

    Application: IF-tissue

    Species: Mouse

    Site: Cerebellum

    Sample: Paraffin-embedded section

    Antibody concentration: 1:500

  • Application: IF-tissue<br /><br />Species: Rat<br /><br />Site: Cerebellum<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1:500

    Application: IF-tissue

    Species: Rat

    Site: Cerebellum

    Sample: Paraffin-embedded section

    Antibody concentration: 1:500

  • Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-EAAT1 antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/8,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-EAAT1 antibody (HA750415) at 1/8,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750415) at 1/8,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-EAAT1 antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-EAAT1 antibody (HA750415) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750415) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-EAAT1 antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-EAAT1 antibody (HA750415) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750415) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of EAAT1 on different lysates with Rabbit anti-EAAT1 antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/5,000 dilution.<br /><br />Lane 1: Mouse brain tissue lysate (no heat)<br />Lane 2: Mouse cerebellum tissue lysate<br />Lane 3: Mouse brain tissue lysate treated with deglycosylation<br />Lane 4: Mouse brain tissue lysate<br />Lane 5: Rat brain tissue lysate (no heat)<br />Lane 6: Rat cerebellum tissue lysate<br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 60 kDa<br />Observed band size: 60-250 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750415" style="font-weight: bold;text-decoration: underline;">HA750415</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of EAAT1 on different lysates with Rabbit anti-EAAT1 antibody (HA750415) at 1/5,000 dilution.

    Lane 1: Mouse brain tissue lysate (no heat)
    Lane 2: Mouse cerebellum tissue lysate
    Lane 3: Mouse brain tissue lysate treated with deglycosylation
    Lane 4: Mouse brain tissue lysate
    Lane 5: Rat brain tissue lysate (no heat)
    Lane 6: Rat cerebellum tissue lysate

    Notice: no heat means the lysate is not boiled.

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 60 kDa
    Observed band size: 60-250 kDa

    Exposure time: 25 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750415) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

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