Pdcd4 (phospho-S457) antibody

• Catalog Number: orb344434
• Category: Antibodies
• Description: Pdcd4 (phospho-S457) antibody
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 9G6
• Tested applications: ELISA, IHC, WB
• Reactivity: Human, Mouse
• Isotype: IgG1
• Immunogen: This monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding Ser457 of the human Pdcd4 protein.
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:20,000 - 1:100,000, IHC: 1:1,000 - 1:5,000, WB: 1:2,000 - 1:10,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This product was purified from concentrated tissue culture supernate by Protein A chromatography. This antibody is specific for human Pdcd4 protein phosphorylated at Ser457. A BLAST analysis was used to suggest cross-reactivity with Pdcd4 from human, mouse, rat and Xenopus based on 100% homology with the immunizing sequence. Cross-reactivity with Pdcd4 from other sources has not been determined.
• Conjugation: Unconjugated
• UniProt ID: Q53EL6
• NCBI: 21735596
• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: mouse anti-Pdcd4 pS457 Antibody, Death up-regulate
• Note: For research use only
• Application notes: Pdcd4 phospho S457 Antibody is tested by ELISA, immunohistochemistry, and western blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 62 kDa in size corresponding to phosphorylated Pdcd4 protein by western blotting in the appropriate cell lysate or extract. This phospho-specific monoclonal antibody reacts with human Pdcd4 pS457 and shows minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide.
• Expiration Date: 12 months from date of receipt.

Biorbyt Antibody orb344433 has been tested in immunohistochemistry, 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. A representative image from positively stained small intestine shows the localization of the anti Pdcd4 antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.

Western blot using Biorbyt 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 at 1:40000 in orb348637 30 min RT. Bands at ~62 kD and ~32 kD were detected.

Western blot using Biorbyt's Protein A purified Mouse Monoclonal anti-Pdcd4 pS457 antibody shows detection of phosphorylated Pdcd4 (indicated by arrowhead at ~62 kDa) in NIH-3T3 cells (p/n orb348723) after 5 min treatment with 30 ng/mL PDGF [lane 2]. No reactivity is seen for (non-phosphorylated) in unstimulated NIH 3T3 cells (p/n orb348714) [lane 1]. The membrane was probed with the primary antibody at a 1:2000 dilution, overnight at 4°C. For detection HRP conjugated Rb-a-Mouse IgG (p/n orb347544) was used at a 1:20000 dilution in blocking buffer (p/n orb348637) for 1 h at 4°C.