Anti-ROBO1 antibody [10E2]

Anti-ROBO1 antibody [10E2]

• 概述

• 产品描述Specific cells in the midline separate the left and right halves of the central nervous system and play many roles in guiding the growth cone behavior. In vertebrate spinal cord, insect abdominal nerves and nematodes, midline cells produce induced cues, such as nectins and slits, respectively, as attractants and repellents. respectively. These cells can serve as gatekeepers to prevent axons from passing through the midline and to guide the growth cone in response to the switching of lead cues beyond the crossing. One such gatherer, Robo, is an axon guidance receptor that defines a new subfamily of proteins from fruit flies to mammalian conserved Ig superfamily proteins. Robo acts as a receptor for the repellent Slit and functions in a cell-autonomous manner Non-intersecting axons express high levels of Robo and cross axons to express low levels of Robo and then reach midline and high levels of crossover. Robo1 and Robo2 are two human homologs of Drosophila megalopolis. Robo1 is also homologous to the C. elegans gene sax3, while Robo2 is homologous to zebrafish genes.
• 产品名称Anti-ROBO1 antibody [10E2]
• 分子量Predicted band size 170-181 kDa.
• 种属反应性Human,Mouse
• 验证应用WB,IHC-P,ICC,FC
• 抗体类型小鼠单抗
• 免疫原Recombinant protein within human ROBO1 aa 40-300.
• 偶联Non-conjugated

• 性能

• 形态Liquid
• 浓度2 mg/ml.
• 存放说明Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
• 存储缓冲液1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
• 亚型IgG1
• 纯化方式Protein G purified.
• 亚细胞定位Membrane.
• 其它名称
  • Deleted in U twenty twenty antibody
  • DUTT 1 antibody
  • DUTT1 antibody
  more

• 应用

  WB: 1:1000-1:5000
  IHC-P: 1:50-1:200
  ICC: 1:50-1:200
  FC: 1:50-1:100

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  Fig1: Western blot analysis of ROBO1 on Siha cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody  was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

   

  

  Fig2: ICC staining of ROBO1 in SHSY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

   

  

  Fig3: ICC staining of ROBO1 in Siha cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody  for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

   

  

  Fig4: Immunohistochemical analysis of paraffin-embedded human Liver cancer tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibodyfor 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig6: Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig7: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-ROBO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibodyfor 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig8: Flow cytometric analysis of ROBO1 was done on Siha cells. The cells were fixed, permeabilized and stained with the primary antibody (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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