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Anti-ZO-1 Antibody

144-61634
RayBiotech
ApplicationsImmunoFluorescence, Western Blot
Product group Antibodies
TargetTJP1
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Overview

  • Supplier
    RayBiotech
  • Product Name
    Anti-ZO-1 Antibody
  • Delivery Days Customer
    16
  • Applications
    ImmunoFluorescence, Western Blot
  • Certification
    Research Use Only
  • Conjugate
    Unconjugated
  • Gene ID7082
  • Target name
    TJP1
  • Target description
    tight junction protein 1
  • Target synonyms
    tight junction protein ZO-1; ZO-1; zona occludens 1; zonula occludens 1 protein
  • Host
    Rabbit
  • Isotype
    IgG
  • Protein IDQ07157
  • Protein Name
    Tight junction protein ZO-1
  • Scientific Description
    ZO-1 pAb
  • Storage Instruction
    -20°C
  • UNSPSC
    12352203

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Immunofluorescence staining of U251 cells with CSB-PA023573HA01HU at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
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Figure 1. Western blot analysis of TJP1 using anti-TJP1 antibody (PB9234). 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 PC-3 whole cell lysates, Lane 2: human CACO-2 whole cell lysates, Lane 3: human COLO320 whole cell lysates, Lane 4: rat testis tissue lysates, Lane 5: mouse testis tissue 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-TJP1 antigen affinity purified polyclonal antibody (Catalog # PB9234) 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 TJP1 at approximately 220 kDa. The expected band size for TJP1 is at 185 kDa.
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293T (Positive) and HL-60 (Negative control) cells (black) were fixed with 4% PFA for 10min at room temperature, permeabilized with PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ZO-1 Antibody( bs-1329R) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) 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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