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Western blot analysis of SMYD3 on different lysates with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/1000 dilution.
Lane 1: Rat kidney tissue lysate
Lane 2: PC-12 cell lysate
Lane 3: Hela cell lysate
Lane 4: Mouse spleen tissue lysate
Lane 5: NIH-3T3 cell lysate
Lane 6: Rat spleen tissue lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 39 kDa
Observed band size: 39 kDa
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-58) at 1/1000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
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"Immunocytochemistry analysis of Hela cells labeling SMYD3 with Rabbit anti-SMYD3 antibody (ET1705-58) 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-SMYD3 antibody (ET1705-58) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI."
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Immunocytochemistry analysis of HepG2 cells labeling SMYD3 with Rabbit anti-SMYD3 antibody (ET1705-58) 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-SMYD3 antibody (ET1705-58) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
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Immunocytochemistry analysis of LOVO cells labeling SMYD3 with Rabbit anti-SMYD3 antibody (ET1705-58) 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-SMYD3 antibody (ET1705-58) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI..
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-SMYD3 antibody. Counter stained with hematoxylin.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.Hela cells
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 (ET1705-58) at 1/50 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.
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Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.Hela cells
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 (ET1705-58) at 1/50 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.
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Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-SMYD3 antibody (ET1705-58) at 1/50 dilution.Hela cells
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 (ET1705-58) at 1/50 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.
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Flow cytometric analysis of Hela cells with SMYD3 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black).
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