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Anti-PKC alpha Antibody

A95177
Antibodies.com
ApplicationsWestern Blot, ELISA, ImmunoHistoChemistry
Product group Antibodies
ReactivityHuman, Mouse, Rat
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Overview

  • Supplier
    Antibodies.com
  • Product Name
    Anti-PKC alpha Antibody
  • Delivery Days Customer
    7
  • Applications
    Western Blot, ELISA, ImmunoHistoChemistry
  • Certification
    Research Use Only
  • Clonality
    Polyclonal
  • Conjugate
    Unconjugated
  • Host
    Rabbit
  • Isotype
    IgG
  • Scientific Description
    Rabbit polyclonal antibody to PKC alpha.
  • Reactivity
    Human, Mouse, Rat
  • UNSPSC
    12352203

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Figure 1. Western blot analysis of PKC Alpha/PRKCA using anti-PKC Alpha/PRKCA antibody (A00743-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Jurkat whole cell lysates, Lane 2: human 293T cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human K562 whole cell lysates, Lane 5: rat C6 whole cell lysates, Lane 6: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-PKC Alpha/PRKCA antigen affinity purified polyclonal antibody (Catalog # A00743-1) 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 1:5000 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 PKC Alpha/PRKCA at approximately 80 kDa. The expected band size for PKC Alpha/PRKCA is at 77 kDa.
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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-PKC alpha/beta II (Thr638/641) 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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