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Western blot analysis of UBE4B on different lysates with Rabbit anti-UBE4B antibody (ET7111-11) at 1/5,000 dilution.
Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: Raji cell lysate (10 µg/Lane)
Lane 3: RAW264.7 cell lysate (10 µg/Lane)
Lane 4: PC-12 cell lysate (10 µg/Lane)
Lane 5: Mouse testis tissue lysate (20 µg/Lane)
Lane 6: Rat testis tissue lysate (20 µg/Lane)
Predicted band size: 146 kDa
Observed band size: 146 kDa
Exposure time: 6 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 (ET7111-11) 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.
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☑ Knockout (KO)
All lanes: Western blot analysis of UBE4B with anti-UBE4B antibody [JE55-76] (ET7111-11) at 1:1,000 dilution.
Lane 1: Wild-type N2A whole cell lysate (30 µg).
Lane 2: UBE4B knockout N2A whole cell lysate (30 µg).
ET7111-11 was shown to specifically react with UBE4B in wild-type N2A cells. No band was observed when UBE4B knockdout sample was tested. Wild-type and UBE4B knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET7111-11, 1:1,000) was used in 5% BSA at room temperature for 2 hours.
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Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7111-11) at 1/200 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 kidney tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 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 (ET7111-11) at 1/200 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 kidney tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 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 (ET7111-11) at 1/200 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 rat kidney tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/200 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 (ET7111-11) at 1/200 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 rat skeletal muscle tissue with Rabbit anti-UBE4B antibody (ET7111-11) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7111-11) 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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