| Rabbit Anti-PDPK1 antibody |
| 产品应用（已验证） |
WB,IHC,ICC,FCM |
| 产品应用（可尝试） |
IP |
| 推荐稀释比例 |
WB=1:500-2000,IP=1:20-100,IHC-P=1:50-200,Flow Cyt=1:50,ICC=1:50, |
| 研究领域 |
细胞生物,神经生物学,信号转导,细胞凋亡,激酶和磷酸酶 |
| 标签 |
Array |
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Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (PDK1) monoclonal Antibody, Unconjugated (bsm-54037R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
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Blank control:NIH/3T3.
Primary Antibody (green line): Rabbit Anti-PDK1 antibody (bsm-54037R)
Dilution: 1:50;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1:1000.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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NIH/3T3 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (PDK1) monoclonal Antibody, Unconjugated (bsm-54037R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
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Paraformaldehyde-fixed, paraffin embedded (human kidney tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PDK1) Monoclonal Antibody, Unconjugated (bsm-54037R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Paraformaldehyde-fixed, paraffin embedded (mouse heart tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PDK1) Monoclonal Antibody, Unconjugated (bsm-54037R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Sample:
Lane 1: rat heart tissue lysate
Lane 2: mouse heart tissue lysate
Primary: Anti-PDK1 (bsm-54037R) at 1:500 dilution
Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution
Predicted band size: 61 kD
Observed band size: 50 kD
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Paraformaldehyde-fixed, paraffin embedded (human myocardium); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PDK1) Monoclonal Antibody, Unconjugated (bsm-54037R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PDK1) Monoclonal Antibody, Unconjugated (bsm-54037R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
RRID：RRID
产品名称：Rabbit Anti-PDPK1 antibody
别名： 3 Phosphoinositide Dependent Protein Kinase 1; hPDK 1; hPDK 1; PDK 1; PkB kinase; PkB kinase like gene 1; PDK1; PkB like 1; PDPK1_HUMAN.
中文名称：PDK1重组兔单克隆抗体
英文名称：Rabbit Anti-PDPK1 antibody
中文别名：3磷酸肌醇依赖性蛋白激酶1
抗体来源： Rabbit
克隆类型：单克隆
细胞定位：细胞浆,细胞膜
性 状：Liquid
亚 型：IgG
纯化方法：affinity purified by Protein A
保存条件：Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
免 疫 原：KLH conjugated synthetic peptide derived from human PDK1
SWISS：O15530
Gene ID ：5170
Human Gene ID：5170
PDK1 (3 Phosphoinositide Dependent Protein Kinase 1) phosphorylates AGC kinases.
PDK1 activates conventional PKC and PKC zeta through phosphorylation of critical threonine residues in the activation loop. PDK1 also phosphorylates Protein Kinase B (PKB) at threonine 308 in the presence of phosphatidylinositol-3,4,5-trisphosphate. Active Akt inactivates Glycogen Synthase Kinase 3 (GSK3), eventually leading to the dephosphorylation and activation of glycogen synthase and the stimulation of glycogen synthesis. Because of the role that PDK plays in insulin-induced glycogen synthesis and PKC activation it is a potentially important target for metabolic drug research. There are three named isoforms.
Function：Phosphorylates and activates not only PKB/AKT, but also PKA, PKC-zeta, RPS6KA1 and RPS6KB1. May play a general role in signaling processes and in development (By similarity). Isoform 3 is catalytically inactive.
Subunit：Interacts with NPRL2.
Subcellular Location：Cytoplasm. Membrane; Peripheral membrane protein. Note=Membrane-associated after cell stimulation leading to its translocation. Tyrosine phosphorylation seems to occur only at the plasma membrane.
Tissue Specificity：Appears to be expressed ubiquitously.
Post-translational modifications：Phosphorylated on tyrosine and serine/threonine. Phosphorylation on Ser-241 in the activation loop is required for full activity. PDK1 itself can autophosphorylate Ser-241, leading to its own activation.
Similarity：Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PDK1 subfamily.
Contains 1 PH domain.
Contains 1 protein kinase domain.
Important Note：This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
400-901-9800
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