製品の概要

  • 製品名
    Anti-Islet 1 antibody [1B1]
    Islet 1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [1B1] to Islet 1
  • 由来種
    Mouse
  • 特異性
    In western blot, we observe a specific band at ~40kDa which is not seen in KO cell lines. A cross-reactive band was observed in the wild-type and knockout cells. The additional band below this band of interest is seen at ~38kDa in both the WT and KO cells. We are unsure as to the identity of these extra bands.
  • アプリケーション
    適用あり: Flow Cyt, IHC-P, WB, ELISA, ICC/IFmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Recombinant fragment corresponding to Human Islet 1 aa 150-349. Purified recombinant fragment expressed in E. Coli.
    Database link: P61371

  • ポジティブ・コントロール
    • WB: Full length Islet 1 (aa 1-349)-hIgGFc transfected HEK293 cell lysate. ICC/IF: HEK293 cells transfected with full length Islet 1-hIgGFc. IHC-P (FFPE): Human Pancreas (Normal) tissue.
  • 特記事項

    This product was changed from ascites to supernatant. Lot no’s high than GR291482-3 are from Tissue Culture Supernatant

     

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab86501 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/500 - 1/2000. Predicted molecular weight: 39 kDa.
ELISA 1/10000.
ICC/IF 1/200 - 1/1000.

ターゲット情報

  • 機能
    Binds to one of the cis-acting domain of the insulin gene enhancer.
  • 組織特異性
    Expressed in subsets of neurons of the adrenal medulla and dorsal root ganglion, inner nuclear and ganglion cell layers in the retina, the pineal and some regions of the brain.
  • 配列類似性
    Contains 1 homeobox DNA-binding domain.
    Contains 2 LIM zinc-binding domains.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Insulin gene enhancer protein ISL 1 antibody
    • Insulin gene enhancer protein ISL-1 antibody
    • Insulin related protein antibody
    • ISL 1 antibody
    • ISL LIM homeobox 1 antibody
    • ISL1 antibody
    • ISL1 transcription factor LIM homeodomain antibody
    • ISL1 transcription factor, LIM/homeodomain (islet 1) antibody
    • ISL1 transcription factor, LIM/homeodomain antibody
    • ISL1_HUMAN antibody
    • Islet-1 antibody
    • Islet1 antibody
    see all

画像

  • All lanes : Anti-Islet 1 antibody [1B1] (ab86501) at 1/500 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Islet 1 knockout HAP1 whole cell lysate
    Lane 3 : SH-SY5Y whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 39 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab86501 observed at 40 kDa. Red - loading control, ab176560, observed at 50 kDa.

    ab86501 was shown to recognize Islet 1 in wild-type HAP1 cells as signal was lost at the expected MW in Islet 1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Islet 1 knockout samples were subjected to SDS-PAGE. ab86501 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Confocal immunofluorescence analysis of HEK293 cells trasfected with full-length Islet 1-hIgGFc using ab86501 at 1/150 dilution (green). Blue: DRAQ5 fluorescent DNA dye.

  • Anti-Islet 1 antibody [1B1] (ab86501) at 1/500 dilution + full length Islet 1-hIgGFc transfected HEK293 cell lysate

    Predicted band size: 39 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    Note: The molecular weight is larger than expected, as the recombinant protein is not the endogenouse protein, but rather the fc-fusion protein.
  • IHC image of Islet 1 staining in Human Pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab86501, 1/1000 dilution, 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.

  • Overlay histogram showing SH-SY5Y cells stained with ab86501 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab86501, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.

参考文献

This product has been referenced in:
  • Burkhardt MF  et al. A cellular model for sporadic ALS using patient-derived induced pluripotent stem cells. Mol Cell Neurosci 56:355-64 (2013). Read more (PubMed: 23891805) »
See 1 Publication for this product

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