ET1704-54

EAAT1 Recombinant Rabbit Monoclonal Antibody [JA30-35]

cat.: ET1704-54

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Tissue, IHC-Fr
Clonality:	Monoclonal
Clone number:	JA30-35

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 60 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human EAAT1 aa 171-220 / 542.
Positive control:	Mouse cerebellum tissue, rat cerebellum tissue, human brain tissue, mouse brain tissue, rat brain tissue, Rat cerebellum tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate.
Subcellular location:	Cell membrane.
Recommended Dilutions: WB IHC-P IF-Tissue IHC-Fr	1:2,000 1:1,000 1:100 1:200
Uniprot #:	SwissProt: P43003 Human \| P56564 Mouse \| P24942 Rat
Alternative names:	GLAST-1 SLC1A3 EAAT1 GLAST GLAST1

Images

	Fig1: Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-EAAT1 antibody (ET1704-54) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-54, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig2: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-EAAT1 antibody (ET1704-54) 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 (ET1704-54) 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.
	Fig3: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-EAAT1 antibody (ET1704-54) 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 (ET1704-54) 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.

	Fig4: Immunofluorescence analysis of paraffin-embedded mouse cerebellum tissue labeling EAAT1 with Rabbit anti-EAAT1 antibody (ET1704-54) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-54, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig5: Immunofluorescence analysis of paraffin-embedded rat cerebellum tissue labeling EAAT1 with Rabbit anti-EAAT1 antibody (ET1704-54) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1704-54, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-EAAT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-54, 1/50) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-EAAT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-54, 1/50) 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.

Fig8: Immunohistochemical analysis of paraffin-embedded human brain tissue using anti-EAAT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1704-54, 1/50) 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.

Fig9: Western blot analysis of EAAT1 on different lysates with Rabbit anti-EAAT1 antibody (ET1704-54) at 1/2,000 dilution.

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

Lysates/proteins at 10 µg/Lane.

Predicted band size: 60 kDa
Observed band size: 60-150 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 (ET1704-54) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.