Validated using a knockout cell line
Recombinant
RabMAb

Anti-Islet 1 antibody [EPR10362] (ab178400)

Overview

  • Product name
    Anti-Islet 1 antibody [EPR10362]
    See all Islet 1 primary antibodies
  • Description
    Rabbit monoclonal [EPR10362] to Islet 1
  • Host species
    Rabbit
  • Specificity
    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.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Human Islet 1 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P61371

  • Positive control
    • HeLa, K562, Jurkat and SH-SY5Y cell lysates; Human gastric neuroendocrine carcinoma and small cell lung carcinoma tissues; SH-SY5Y cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab178400 in the following tested applications.

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

Application Abreviews Notes
ICC/IF 1/250 - 1/500.
Flow Cyt Use at an assay dependent concentration.
IP 1/10 - 1/100.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/1000 - 1/10000. Predicted molecular weight: 39 kDa.

Target

  • Function
    Binds to one of the cis-acting domain of the insulin gene enhancer.
  • Tissue specificity
    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.
  • Sequence similarities
    Contains 1 homeobox DNA-binding domain.
    Contains 2 LIM zinc-binding domains.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • Lanes 1-3 : Anti-Islet 1 antibody [EPR10362] (ab178400) at 1/1000 dilution
    Lane 4 : Anti-Islet 1 antibody [EPR10362] (ab178400) at 1000 µg

    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



    Lanes 1 - 4: Merged signal (red and green). Green - ab178400 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab178400 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. Ab178400 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling Islet 1 with purified ab178400 at 1:200 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1:2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human gastric neuroendocrine carcinoma tissue labeling Islet 1 with ab178400 at a 1/250 dilution.

  • All lanes : Anti-Islet 1 antibody [EPR10362] (ab178400) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : K562 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Developed using the ECL technique.

    Predicted band size: 39 kDa

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human small cell lung carcinoma tissue labeling Islet 1 with ab178400 at a 1/250 dilution.

  • Immunofluorescence analysis of SH-SY5Y cells labeling Islet 1 with ab178400 at a 1/250 dilution.

  • Western blot analysis on Immunoprecipitation pellet from either 1) SH-SY5Y cell lysate, or 2) 1xPBS (negative control); showing Islet 1, using ab178400 at 1/10 dilution and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

References

This product has been referenced in:
  • Xiang Q  et al. ISL1 overexpression enhances the survival of transplanted human mesenchymal stem cells in a murine myocardial infarction model. Stem Cell Res Ther 9:51 (2018). Read more (PubMed: 29482621) »

See 1 Publication for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (Human pluripotent stem cell derived cardiomyocyte)
Specification
Human pluripotent stem cell derived cardiomyocyte
Permeabilization
Yes - saponin
Fixative
Paraformaldehyde
Username

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Verified customer

Submitted Oct 16 2014

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