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  1. Link

    products/primary-antibodies/islet-1-antibody-epr10362-ab178400.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Islet 1 antibody [EPR10362] (ab178400)

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  • SDS
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)
  • Anti-Islet 1 antibody [EPR10362] (ab178400)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10362] to Islet 1
  • Suitable for: IP, IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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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: IP, IHC-P, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • 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

    • WB: HeLa, K562, A431, A375, Jurkat and SH-SY5Y cell lysates. IHC-P: Human gastric neuroendocrine carcinoma, pancreas and small cell lung carcinoma tissues. IP: SH-SY5Y cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR10362
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Stem Cells
    • Lineage Markers
    • Endoderm
    • Stem Cells
    • Neural Stem Cells
    • Intracellular
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Cardiovascular
    • Heart
    • Cardiogenesis
    • Transcription factors/regulators
    • Stem Cells
    • Neural Stem Cells
    • Neuron Restricted Lineage
    • Developmental Biology
    • Lineage specification
    • Endoderm
    • Developmental Biology
    • Organogenesis
    • Gut development
    • Pancreas development
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (ab238919)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab7899)
    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant Human Islet 1 protein (ab134882)
  • Related Products

    • Recombinant Human Islet 1 protein (ab134882)

Applications

The Abpromise guarantee

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
IP
1/10 - 1/100.
IHC-P
1/6000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500

WB
1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
Notes
IP
1/10 - 1/100.
IHC-P
1/6000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500

WB
1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

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.
  • Target information above from: UniProt accession P61371 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3670 Human
    • Omim: 600366 Human
    • SwissProt: P61371 Human
    • Unigene: 505 Human
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreas tissue sections labeling Islet 1 with purified ab178400 at 1/6000 dilution (0.03 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
      All lanes : Anti-Islet 1 antibody [EPR10362] (ab178400) at 1/1000 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



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

      Unpurified 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 1/1,000 dilution and 1/20,000 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 1/20,000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
      All lanes : Anti-Islet 1 antibody [EPR10362] (ab178400) at 1/10000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 39 kDa
      Observed band size: 39 kDa

    • Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)

      ab178400 (purified ) at 1/20 dilution (1ug) immunoprecipitating Islet 1 in SH-SY5Y whole cell lysate.
      Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug
      Lane 2 (+): ab178400 & SH-SY5Y whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab178400 in SH-SY5Y whole cell lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)

      Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Islet 1 with ab178400  followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on human pancreatic islet. The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    • Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Western blot - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Anti-Islet 1 antibody [EPR10362] (ab178400) at 1/10000 dilution ((unpurified)) + A375 (HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 39 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)

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

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)

      Islet 1 was immunoprecipitated from 0.35 mg SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10μg with unpurifed ab178400 at 1:50 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using unpurified ab178400 1:1000 dilution (2 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.

      Lane 1: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10μg

      Lane 2: ab178400 IP in SH-SY5Y whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab178400 in SH-SY5Y whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 1 second

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

      Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : K562 (human chronic myelogenous leukemia cell line from bone marrow ) cell lysate
      Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
      Lane 4 : SH-SY5Y (human neuroblastoma cell line from bone marrow) 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [EPR10362] (ab178400)

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

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)
      Immunoprecipitation - Anti-Islet 1 antibody [EPR10362] (ab178400)

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

    • Anti-Islet 1 antibody [EPR10362] (ab178400)
      Anti-Islet 1 antibody [EPR10362] (ab178400)

    Protocols

    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab178400? Please let us know so that we can cite the reference in this datasheet.

    ab178400 has been referenced in 2 publications.

    • Jin X  et al. ERa is required for suppressing OCT4-induced proliferation of breast cancer cells via DNMT1/ISL1/ERK axis. Cell Prolif 52:e12612 (2019). PubMed: 31012189
    • 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). PubMed: 29482621

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Islet 1 antibody [EPR10362]

    Excellent
    Abreviews
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 16 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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