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SMAD1 antibody

10R-1435
Biosynth
Product group Antibodies
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Overview

  • Supplier
    Biosynth
  • Product Name
    SMAD1 antibody
  • Delivery Days Customer
    7
  • Certification
    Research Use Only
  • UNSPSC
    12352203

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Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃,and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad1 + Smad5 Antibody(bs-1619R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
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Figure 1. Western blot analysis of SMAD1 using anti-SMAD1 antibody (A00728-1). 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 50ug of sample under reducing conditions. Lane 1: rat kidney tissue lysate, Lane 2: rat testis tissue lysate, Lane 3: mouse brain tissue lysate, Lane 4: mouse kidney tissue lysate, Lane 5: mouse testis tissue lysate, Lane 6: human Hela cell lysate, Lane 7: human A375 cell lysate, Lane 8: human HepG2 cell lysate. 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-SMAD1 antigen affinity purified polyclonal antibody (Catalog # A00728-1) 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 1:10000 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 SMAD1 at approximately 52,60KD. The expected band size for SMAD1 is at 48, 52, 60KD.
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