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Western blot analysis of VDAC1 on different lysates with Rabbit anti-VDAC1 antibody (R1307-1) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: Raji cell lysate
Lane 5: HepG2 cell lysate
Lane 6: SW480 cell lysate
Lane 7: A549 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 31 kDa
Observed band size: 31 kDa
Exposure time: 1 minute 30 seconds;
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 (R1307-1) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of VDAC1 on different lysates with Rabbit anti-VDAC1 antibody (R1307-1) at 1/2,000 dilution.
Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: C2C12 cell lysate (20 µg/Lane)
Lane 3: C6 cell lysate (20 µg/Lane)
Lane 4: Mouse skeletal muscle tissue lysate (40 µg/Lane)
Lane 5: Rat skeletal muscle tissue lysate (40 µg/Lane)
Predicted band size: 31 kDa
Observed band size: 31 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 (R1307-1) at 1/2,000 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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☑ Knockdown (KD)
Western blot analysis of VDAC1 on different lysates with Rabbit anti-VDAC1 antibody (R1307-1) at 1/1,000 dilution.
Lane 1: HEK293-si NT cell lysate (10 µg/Lane)
Lane 2: HEK293-si VDAC1#1(no heat) cell lysate (10 µg/Lane)
Lane 3: HEK293-si VDAC1#2(no heat) cell lysate (10 µg/Lane)
Notice: no heat means the lysate is not boiled.
Predicted band size: 31 kDa
Observed band size: 31 kDa
Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
R1307-1 was shown to specifically react with VDAC1 in HEK293-si NT cells. No band were observed when HEK293-si VDAC1 samples were tested. HEK293-si NT and HEK293-si VDAC1 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 (R1307-1, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at 4℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
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ICC staining VDAC1 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (blue).
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Immunohistochemical analysis of paraffin-embedded human skeletal mucle tissue with Rabbit anti-VDAC1 antibody (R1307-1) at 1/500 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 (R1307-1) at 1/500 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 skeletal mucle tissue with Rabbit anti-VDAC1 antibody (R1307-1) at 1/500 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 (R1307-1) at 1/500 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 mucle tissue with Rabbit anti-VDAC1 antibody (R1307-1) at 1/500 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 (R1307-1) at 1/500 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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