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Western blot analysis of Phospho-PDHA1 (S293) on different lysates with Rabbit anti-Phospho-PDHA1 (S293) antibody (HA721443) at 1/1,000 dilution.
Lane 1: A549 cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: HeLa cell lysate (20 µg/Lane)
Lane 5: HEK-293 cell lysate (20 µg/Lane)
Lane 6: Human kidney tissue lysate (40 µg/Lane)
Lane 7: Mouse brain tissue lysate (40 µg/Lane)
Lane 8: Mouse heart tissue lysate (40 µg/Lane)
Predicted band size: 43 kDa
Observed band size: 43 kDa
Exposure time: 39 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 (HA721443) 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 Phospho-PDHA1 (S293) on different lysates with Rabbit anti-Phospho-PDHA1 (S293) antibody (HA721443) at 1/1,000 dilution.
Lane 1: Rat kidney tissue lysate
Lane 2: Mouse kidney tissue lysate
Lysates/proteins at 30 µg/Lane.
Predicted band size: 43 kDa
Observed band size: 43 kDa
Exposure time: 10 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 (HA721443) 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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Immunocytochemistry analysis of Jurkat cells treated with or without λpp labeling Phospho-PDHA1 (S293) with Rabbit anti-Phospho-PDHA1 (S293) antibody (HA721443) 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-Phospho-PDHA1 (S293) antibody (HA721443) at 1/100 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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Immunohistochemical analysis of paraffin-embedded human renal clear cell carcinoma tissue with Rabbit anti-Phospho-PDHA1 (S293) antibody (HA721443) 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 (HA721443) 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 brain tissue with Rabbit anti-Phospho-PDHA1 (S293) antibody (HA721443) 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 (HA721443) 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 colon tissue with Rabbit anti-Phospho-PDHA1 (S293) antibody (HA721443) at 1/2,000 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 (HA721443) at 1/2,000 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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