EAAT3 Recombinant Rabbit Monoclonal Antibody [JU39-69]
cat.: ET1706-55
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Rat, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JU39-69
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 57 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human EAAT3 aa 80-209 / 524.
Positive control: Human brain tissue lysate, Human lung tissue lysate, SH-SY5Y, rat brain tissue, human tonsil tissue, human colon carcinoma tissue, human kidney tissue.
Subcellular location: Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:1,000
Uniprot #: SwissProt: P43005 Human | P51906 Mouse | P51907 Rat
Alternative names: EAA3_HUMAN EAAC 1 EAAC1 EAAT 3 Excitatory amino acid carrier 1 Excitatory amino acid carrier1 Excitatory amino acid transporter 3 Excitatory amino acid transporter3 Excitatory amino-acid carrier 1 GLUTAMATE TRANSPORTER, HIGH-AFFINITY MEAAC 1 MEAAC1 Neuronal and epithelial glutamate transporter REAAC 1 REAAC1 Slc1 a1 Slc1a 1 SLC1A1 Sodium dependent glutamate/aspartate transporter 3 Sodium-dependent glutamate/aspartate transporter 3 Solute carrier family 1 (neuronal / epithelial high affinity glutamate transporter, system Xag), member 1 SOLUTE CARRIER FAMILY 1 (NEURONAL/EPITHELIAL HIGH AFFINITY GLUTAMATE TRANSPORTER), MEMBER 1 Solute carrier family 1 (neuronal/epithelial high affinity glutamate transporter, system Xag), member 1 Solute carrier family 1 member 1 Solute carrier family 1, member 1
Images
ET1706-55_1.jpg Fig1: Western blot analysis of EAAT3 on different lysates with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/2,000 dilution.

Lane 1: Human brain tissue lysate
Lane 2: Human lung tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 60 kDa

Exposure time: 30 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 (ET1706-55) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1706-55_2.jpg Fig2: Immunocytochemistry analysis of SH-SY5Y cells labeling EAAT3 with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-55_3.jpg Fig3: Flow cytometric analysis of SH-SY5Y cells labeling EAAT3.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1706-55, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1706-55_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/50 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 (ET1706-55) at 1/50 dilution for 0.5 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.
ET1706-55_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/50 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 (ET1706-55) at 1/50 dilution for 0.5 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.
ET1706-55_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/50 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 (ET1706-55) at 1/50 dilution for 0.5 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.
ET1706-55_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-EAAT3 antibody (ET1706-55) at 1/50 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 (ET1706-55) at 1/50 dilution for 0.5 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.