Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Islet 1 antibody [EPR10362] - BSA and Azide free (ab238919)

Overview

  • Product name

    Anti-Islet 1 antibody [EPR10362] - BSA and Azide free
    See all Islet 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR10362] to Islet 1 - BSA and Azide free
  • 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: WB, IHC-P, IPmore 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

    • 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

    Ab238919 is the carrier-free version of ab178400. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab238919 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab238919 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
WB Use at an assay dependent concentration. Predicted molecular weight: 39 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400)
  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded, Human gastric neuroendocrine carcinoma tissue labeling Islet 1 with ab178400 at a 1/250 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178400).

References

ab238919 has not yet been referenced specifically in any publications.

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