使用敲除细胞株进行验证

Anti-Integrin alpha 5抗体(ab112183)

概述

  • 产品名称
    Anti-Integrin alpha 5抗体
    参阅全部 Integrin alpha 5 一抗
  • 描述
    兔多克隆抗体to Integrin alpha 5
  • 经测试应用
    适用于: WB, IHC-Pmore details
  • 种属反应性
    与反应: Human
    预测可用于: Cow, Pig, Chimpanzee, Macaque monkey, Gorilla, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 600 - 700 of Human Integrin alpha 5.

  • 阳性对照
    • WB: This antibody gave a positive signal in Human Placenta tissue lysate, JEG-3 whole cell lysate, HeLa whole cell lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal placenta.

性能

应用

Our Abpromise guarantee covers the use of ab112183 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 145 kDa (predicted molecular weight: 114 kDa).
IHC-P Use a concentration of 1 µg/ml.

靶标

  • 功能
    Integrin alpha-5/beta-1 is a receptor for fibronectin and fibrinogen. It recognizes the sequence R-G-D in its ligands. In case of HIV-1 infection, the interaction with extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions.
  • 序列相似性
    Belongs to the integrin alpha chain family.
    Contains 7 FG-GAP repeats.
  • 细胞定位
    Membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • CD49 antigen-like family member E antibody
    • CD49e antibody
    • Fibronectin receptor subunit alpha antibody
    • Fibronectin receptor, alpha subunit antibody
    • FNRA antibody
    • Integrin alpha 5 (fibronectin receptor alpha) antibody
    • Integrin alpha-5 antibody
    • Integrin alpha-5 light chain antibody
    • Integrin alpha-F antibody
    • Integrin, alpha 5 (fibronectin receptor, alpha polypeptide) antibody
    • ITA5_HUMAN antibody
    • Itga5 antibody
    • Very late activation protein 5, alpha subunit antibody
    • VLA-5 antibody
    • VLA5 antibody
    • VLA5A antibody
    see all

图片



  • Predicted band size : 114 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Integrin alpha 5 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Jeg3 whole cell lysate (20 µg)
    Lane 4: HeLa whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab112183 observed at 140 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab112183 was shown to recognize Integrin alpha 5 when Integrin alpha 5 knockout samples were used, along with additional cross-reactive bands. Wild-type and Integrin alpha 5 knockout samples were subjected to SDS-PAGE. Ab112183 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

     

  • IHC image of Integrin alpha 5 staining in Human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab112183, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Integrin alpha 5 antibody (ab112183) at 1 µg/ml

    Lane 1 : Human placenta tissue lysate - total protein (ab29745)
    Lane 2 : JEG-3 (Human placental choriocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 114 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 32 kDa,50 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 secondsIntegrin alpha 5 contains an extensive number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

文献

ab112183 has not yet been referenced specifically in any publications.

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