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Anti-PDCD4 PHOSPHO Antibody 9G6
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Also for PDCD4 (NM_145341)
|This monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding Ser457 of the human Pdcd4 protein.|
|human, mouse, rat, xenopus
||ELISA: 1:20,000 - 1:100,000, WB: 1:2,000 - 1:10,000, IHC: 1:1,000 - 1:5,000
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|Programmed cell death 4 (Pdcd4) is a novel tumor supressor. Pdcd4 directly inhibits the helicase activity of eukaryotic translation initiation factor 4A (eIF4A), a component of the translation initiation complex. Pdcd4 also suppresses the transactivation of activator protein-1 (AP-1)-responsive promoters by c-Jun. Pdcd4 contains two Akt phosphoryl-ation sites, one at Ser67 and the other at Ser457. The phosphorylation of Pdcd4 by Akt causes nuclear translocation of Pdcd4 and a significant decrease in the ability of Pdcd4 to interfere with the transactivation of AP-1-responsive promoters by c-Jun.
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WB using anti-Pdcd4 pS457 antibody shows detection of phosphorylated Pdcd4 (indicated by arrowhead at ~62 kDa) in NIH-3T3 cells after 5 min treatment with 30 ng/mL PDGF (lane 2). No reactivity is seen for unstimulated (non-phosphorylated) NIH 3T3 cells (lane 1). The membrane was probed with the primary antibody at a 1:2,000 dilution, overnight at 4?. For detection HRP conjugated Rb-a-Mouse IgG was used at a 1:20,000 dilution in blocking buffer for 1 h at 4?.
Western blot using Immunochemicals Protein A purified Mouse Monoclonal anti-Pdcd4 pS457 antibody against recombinant PDCD4 protein. Membrane was blocked in 1% BSA-TBS-T for 30 min RT and probed with 1° Ab Ms-A-Pdcd4pS457 1:1000 (o/n 4°C in 1% BSA-TBS-T) followed by 2° Ab Peroxidase Conjugated Rabbit anti Ms CUST10M Lot# 20121 at 1:40,000 in MB-070 30 min RT. Bands at ~62 kD and ~32 kD were detected.
Antibody TA319574 has been tested in IHC, analyzed by an anatomic pathologist and validated for use in IHC applications against formalin-fixed, paraffin-embedded human tissues. The antibody was serially diluted and tested at a range of concentrations on at least 22 different human formalin-fixed, paraffin archival tissues, and positive and negative tissues were scored and compared to the published literature on the expression and function of the gene.