bsm-54037R [Primary Antibody]
Rabbit  Anti-PDPK1 Recombinant Rabbit mAb   Antibody
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Host: Rabbit

Target Protein: PDPK1 Recombinant Rabbit mAb

IR: Immunogen Range:

Clonality:

Isotype: IgG

Entrez Gene: 5170

Swiss Prot: O15530

Source: KLH conjugated synthetic peptide derived from human PDK1: 

Purification: affinity purified by Protein A

Storage: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

Background: 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.

Size: 50ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:50-200,IHC-F=1:400-800,IF=1:100-500,Flow-Cyt=1:50

Cross Reactive Species: Human,Mouse,Rat

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
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
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.
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.
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.
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.
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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