ATP5O Recombinant Rabbit Monoclonal Antibody [PSH15-04]
cat.: HA723704
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, IF-Tissue, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH15-04 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 23 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human ATP5O aa 1-213. |
| Positive control: | K-562 cell lysate, HeLa cell lysate, HepG2 cell lysate, A549 cell lysate, Neuro-2a cell lysate, A20 cell lysate, PC-12 cell lysate, HeLa, Neuro-2a, PC-12, human kidney tissue, human testis tissue, mouse kidney tissue, mouse testis tissue, rat kidney tissue, rat testis tissue. |
| Subcellular location: | Mitochondrion, Mitochondrion inner membrane. |
| Recommended Dilutions:
WB IF-Cell IHC-P IF-Tissue FC IP |
1:5,000 1:100 1:500-1:2,000 1:200-1:500 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P48047 Human | Q9DB20 Mouse | Q06647 Rat |
| Alternative names: | ATP synthase O subunit mitochondrial precursor ATP synthase subunit O ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit ATP5O ATPO ATPO_HUMAN mitochondrial Mitochondrial ATP synthase, O subunit Oligomycin sensitivity conferral protein OSCP |
Images
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Fig1:
Western blot analysis of ATP5O on different lysates with Rabbit anti-ATP5O antibody (HA723704) at 1/5,000 dilution. Lane 1: K-562 cell lysate Lane 2: HeLa cell lysate Lane 3: HepG2 cell lysate Lane 4: A549 cell lysate Lane 5: Neuro-2a cell lysate Lane 6: A20 cell lysate Lane 7: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 23 kDa Observed band size: 23 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 (HA723704) 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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Fig2:
Immunocytochemistry analysis of HeLa cells labeling ATP5O with Rabbit anti-ATP5O antibody (HA723704) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP5O antibody (HA723704) at 1/100 dilution in 1% BSA in PBST 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI. |
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Fig3:
Immunocytochemistry analysis of Neuro-2a cells labeling ATP5O with Rabbit anti-ATP5O antibody (HA723704) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP5O antibody (HA723704) at 1/100 dilution in 1% BSA in PBST 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI. |
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Fig4:
Immunocytochemistry analysis of PC-12 cells labeling ATP5O with Rabbit anti-ATP5O antibody (HA723704) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATP5O antibody (HA723704) at 1/100 dilution in 1% BSA in PBST 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. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ATP5O antibody (HA723704) 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 (HA723704) 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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Fig6:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-ATP5O antibody (HA723704) 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 (HA723704) 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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Fig7:
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ATP5O antibody (HA723704) 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 (HA723704) 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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Fig8:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-ATP5O antibody (HA723704) 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 (HA723704) 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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Fig9:
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-ATP5O antibody (HA723704) 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 (HA723704) 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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Fig10:
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-ATP5O antibody (HA723704) 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 (HA723704) 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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Fig11:
Flow cytometric analysis of HeLa cells labeling ATP5O. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723704, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig12:
Flow cytometric analysis of PC-12 cells labeling ATP5O. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723704, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig13:
ATP5O was immunoprecipitated from 0.2 mg HeLa cell lysate with HA723704 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723704 at 1/5,000 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: HeLa cell lysate (input) Lane 2: HA723704 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA723704 in HeLa cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 3 minutes; ECL: K1801 |
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