Eph receptor B3 Recombinant Rabbit Monoclonal Antibody [JE54-73]
cat.: ET7110-80
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE54-73
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 110 kDa.
Isotype: IgG
Immunogen: Recombinant protein within Human Eph receptor B3 aa 80-220 / 998.
Positive control: Rat liver tissue lysates, MCF-7, RWPE-1, rat kidney tissue, human skin tissue, human prostate carcinoma tissue, MCF-7.
Subcellular location: Cell membrane, dendrite.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:2,000
1:50-1:100
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P54753 Human
Alternative names: Cek10 EK2 Embryonic kinase 2 EPH Like Tyrosine Kinase 2 EPH receptor B3 EPH-like kinase 2 ephb3 EPHB3_HUMAN Ephrin receptor EphB3 Ephrin type B receptor 3 Ephrin type-B receptor 3 ETK2 hEK2 Human Embryo Kinase 2 Mdk5 Sek4 TYRO6 Tyrosine protein kinase receptor HEK2 Tyrosine protein kinase TYRO6 Tyrosine-protein kinase TYRO6
Images
ET7110-80_1.jpg Fig1: Western blot analysis of Eph receptor B3 on rat liver tissue lysates with Rabbit anti-Eph receptor B3 antibody (ET7110-80) at 1/500 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 110 kDa
Observed band size: 110 kDa

Exposure time: 2 minutes;

10% 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 (ET7110-80) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET7110-80_2.jpg Fig2: Immunocytochemistry analysis of MCF-7 cells labeling Eph receptor B3 with Rabbit anti-Eph receptor B3 antibody (ET7110-80) at 1/200 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-Eph receptor B3 antibody (ET7110-80) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET7110-80_3.jpg Fig3: Immunocytochemistry analysis of RWPE-1 cells labeling Eph receptor B3 with Rabbit anti-Eph receptor B3 antibody (ET7110-80) at 1/100 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-Eph receptor B3 antibody (ET7110-80) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET7110-80_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Eph receptor B3 antibody (ET7110-80) at 1/200 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-80) at 1/200 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.
ET7110-80_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Eph receptor B3 antibody (ET7110-80) at 1/200 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-80) at 1/200 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.
ET7110-80_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-Eph receptor B3 antibody (ET7110-80) at 1/200 dilution.

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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-80) at 1/200 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.
ET7110-80_7.jpg Fig7: Flow cytometric analysis of Eph receptor B3 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-80, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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