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Neuroscience Cell Type Marker Neuron marker Soma marker

Anti-Islet 1 antibody [1B1] (ab86501)

Price and availability

284 121 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-Islet 1 antibody [1B1] (ab86501)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Mouse monoclonal [1B1] to Islet 1
  • Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
  • Knockout validated
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG1

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Overview

  • Product name

    Anti-Islet 1 antibody [1B1]
    See all Islet 1 primary antibodies
  • Description

    Mouse monoclonal [1B1] to Islet 1
  • Host species

    Mouse
  • 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 & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    Recombinant fragment
    See all applications and species data
  • Immunogen

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

  • Positive control

    • 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.
  • General notes

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

     

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    Purified from tissue culture supernatant.
  • Clonality

    Monoclonal
  • Clone number

    1B1
  • Isotype

    IgG1
  • 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

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [1B1] (ab86501)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody [1B1] (ab86501)

    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.

  • Western blot - Anti-Islet 1 antibody [1B1] (ab86501)
    Western blot - Anti-Islet 1 antibody [1B1] (ab86501)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Islet 1 antibody [1B1] (ab86501)
    Immunocytochemistry/ Immunofluorescence - Anti-Islet 1 antibody [1B1] (ab86501)

    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.

  • Western blot - Anti-Islet 1 antibody [1B1] (ab86501)
    Western blot - Anti-Islet 1 antibody [1B1] (ab86501)
    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.
  • Flow Cytometry - Anti-Islet 1 antibody [1B1] (ab86501)
    Flow Cytometry - Anti-Islet 1 antibody [1B1] (ab86501)
    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.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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