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Anti-DKK1 Antibody

130-10224
RayBiotech
ApplicationsWestern Blot, ELISA
Product group Antibodies
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Overview

  • Supplier
    RayBiotech
  • Product Name
    Anti-DKK1 Antibody
  • Delivery Days Customer
    16
  • Applications
    Western Blot, ELISA
  • Certification
    Research Use Only
  • Clonality
    Monoclonal
  • Host
    Mouse
  • Scientific Description
    Mouse anti-DKK1(5D2-F6-E4) Antibody
  • Storage Instruction
    -20°C
  • UNSPSC
    12352203

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Figure 1. Western blot analysis of DKK1 using anti-DKK1 antibody (A00632). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. Lane 1: recombinant human DKK1 protein 1ng. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DKK1 antigen affinity purified polyclonal antibody (Catalog # A00632) at 0.5 microg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for DKK1 at approximately 38KD. The expected band size for DKK1 is at 26KD.
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Image kindly provided by Dr. Magdalena Krol. Control tumor cells, tumor cells grown in macrophage-conditioned medium, tumor cells sorted from co-culture with macrophages, and macrophages from monocultures and sorted from co-culture with tumor cells were analyzed. Total protein concentrations in lysates were determined using a Bio-Rad protein assay. Proteins (50 mg) were resolved using SDS-PAGE and transferred onto PVDF membranes. The membranes were then blocked with 5% non-fat dry milk in TBS buffer containing 0.5% Tween 20. The membranes were then incubated overnight with the primary Rabbit Anti-DKK1 Polyclonal Antibody at 1:100 dilution. Subsequently, the membranes were washed three times in TBS containing 0.5% Tween 20 and incubated for 1 h at room temperature with secondary antibodies conjugated with the appropriate infrared (IR) fluorophore IRDyeH 800 CW or IRDyeH 680 RD at a dilution of 1:5000.
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