PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30]
cat.: ET1704-66
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IP, FC, IF-Cell, IF-Tissue |
| Clonality: | Monoclonal |
| Clone number: | JA67-30 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 49 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human PDK1 aa 16-65 / 436. |
| Positive control: | Mouse heart tissue lysate, rat heart tissue lysate, Hela, NIH/3T3, mouse heart tissue, rat heart tissue. |
| Subcellular location: | Mitochondrion matrix. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC IF-Tissue |
1:5,000 1:50-1:200 1:2,000 1:50-1:100 1:200-1:400 |
| Uniprot #: | SwissProt: Q15118 Human | Q8BFP9 Mouse | Q63065 Rat |
| Alternative names: | [Pyruvate dehydrogenase [lipoamide]] kinase isozyme 1, mitochondrial HGNC:8809 Mitochondrial pyruvate dehydrogenase kinase isoenzyme 1 PDH kinase 1 Pdk1 PDK1_HUMAN Pyruvate dehydrogenase kinase isoform 1 Pyruvate dehydrogenase kinase, isoenzyme 1 |
Images
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Fig1:
Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (ET1704-66) at 1/5,000 dilution. Lane 1: Mouse heart tissue lysate Lane 2: Rat heart tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 49 kDa Observed band size: 49 kDa Exposure time: 10 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 (ET1704-66) at 1/5,000 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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Fig2: ICC staining of PDK1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
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Fig3: ICC staining of PDK1 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-PDK1 antibody (ET1704-66) 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 (ET1704-66) 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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Fig5:
Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-PDK1 antibody (ET1704-66) 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 (ET1704-66) 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: Flow cytometric analysis of PDK1 was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-66, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig7:
PDK1 was immunoprecipitated from 0.2 mg mouse heart tissue lysate with ET1704-66 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1704-66 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Mouse heart tissue lysate (input) Lane 2: ET1704-66 IP in mouse heart tissue lysate Lane 3: Rabbit IgG instead of ET1704-66 in mouse heart tissue lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.