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☑ Knockdown (KD)
Western blot analysis of Glycogen synthase 1 / GYS1 on different lysates with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Glycogen synthase 1 / GYS1 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 81 kDa
Observed band size: 81 kDa
Exposure time: 60 seconds; ECL: K1802;
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 (ET1611-59) at 1/1,000 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.
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Western blot analysis of Glycogen synthase 1 / GYS1 on different lysates with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) at 1/2,000 dilution.
Lane 1: HEK-293 cell lysate
Lane 2: A431 cell lysate
Lane 3: RD cell lysate
Lane 4: Mouse heart tissue lysate
Lane 5: Mouse skeletal muscle tissue lysate
Lane 6: Rat heart tissue lysate
Lane 7: Rat skeletal muscle tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 84 kDa
Observed band size: 84 kDa
Exposure time: Lane 1-3: 46 seconds; Lane 4-7: 8 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 (ET1611-59) 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.
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Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) 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 (ET1611-59) 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.
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Immunohistochemical analysis of paraffin-embedded mouse cardiac muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) 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 (ET1611-59) 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.
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Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue with Rabbit anti-Glycogen synthase 1 / GYS1 antibody (ET1611-59) 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 (ET1611-59) 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.
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Glycogen synthase 1 / GYS1 was immunoprecipitated from 0.2 mg A431 cell lysate with ET1611-59 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1611-59 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: A431 cell lysate (input)
Lane 2: ET1611-59 IP in A431 cell lysate
Lane 3: Rabbit IgG instead of ET1611-59 in A431 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801
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