Recombinant
RabMAb

Recombinant Anti-Islet 1 antibody [EP4182] (ab109517)

Rabbit recombinant monoclonal Islet 1 antibody [EP4182]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Chicken, Human. Cited in 18 publication(s). Independently reviewed in 4 review(s).

Overview

  • Product name
    Anti-Islet 1 antibody [EP4182]
    See all Islet 1 primary antibodies
  • Description
    Rabbit monoclonal [EP4182] to Islet 1
  • Host species
    Rabbit
  • Specificity
    86% identities with Islet 2
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, WB, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Chicken, Human
  • Immunogen

    Synthetic peptide within Human Islet 1 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: P61371

  • Positive control
    • WB: K562, HeLa, Jurkat and SH-SY5Y cell lysates. IHC-Fr: Chicken hindbrain tissue. ICC/IF: U-87 MG cell line.
  • General notes

    Rat: We have preliminary testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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 ab109517 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
IHC-Fr 1/1000.
ICC/IF 1/250.

For unpurified use at 1/50 - 1/100.

WB 1/10000 - 1/50000. Predicted molecular weight: 39 kDa.
IP 1/10 - 1/100.
Flow Cyt Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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

  • For immunofluorescence, tissues were collected from mice, embedded fresh into OCT, and subsequently cut into 10μm sections using a Leica Cryostat. Embryos collected after lentiviral infection for the Shh overexpression experiment were pre-fixed for in 4% PFA 1h at RT. Slides were fixed for 10 min (or 7 min for slides of Shh overexpression embryos) in 4% PFA and blocked for 1h or overnight in PBS-Triton with BSA/NDS. Primary antibodies were diluted in blocking solution and incubations were carried out for 1h or overnight, followed by incubation in secondary antibodies for 1h at room temperature. Slides were then counterstained with DAPI and mounted using antifade mounting media.

  • Immunocytochemistry/Immunofluorescence analysis of SH-SY-5Y (human neuroblastoma) cells labelling Islet 1 with purified ab109517 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • All lanes : Anti-Islet 1 antibody [EP4182] (ab109517) at 1/10000 dilution (unpurified)

    Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) 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.

    Predicted band size: 39 kDa

  • Anti-Islet 1 antibody [EP4182] (ab109517) at 1/50000 dilution (purified) + RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 39 kDa



    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

  • Anti-Islet 1 antibody [EP4182] (ab109517) at 1/10000 dilution (purified) + UMNSAH/DF-1 cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 39 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemistry/Immunofluorescence analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Islet 1 with unpurified ab109517.

  • Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling Islet 1 with purified ab109517 at 1/50 dilution(10 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • IHC-Fr image of anti-Islet 1 staining with unpurified ab109517 on tissue sections from chicken hindbrain. The sections were blocked with 3% BSA for 1 hour at 4°C, before incubation with unpurified ab109517 (1/1000 dilution) for 16 hours at 4°C. The secondary was an Alexa-Fluor® 568 conjugated goat anti-rabbit polyclonal, used at a 1/1000 dilution.

    See Abreview

  • Anti-Islet 1 antibody [EP4182] (ab109517) at 1/50000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 39 kDa



    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

  • Anti-Islet 1 antibody [EP4182] (ab109517) at 1/50000 dilution (purified) + K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 39 kDa



    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

  • ab109517 (purified) at 1/30 immunoprecipitating Islet 1 in Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate. 10 µg of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1,500) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

  • ab109517 (unpurified) immunoprecipitating Islet 1 in Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate.

References

This product has been referenced in:
  • Lim GS  et al. Microhexagon gradient array directs spatial diversification of spinal motor neurons. Theranostics 9:311-323 (2019). Read more (PubMed: 30809276) »
  • Zhang Q  et al. Collaborative ISL1/GATA3 interaction in controlling neuroblastoma oncogenic pathways overlapping with but distinct from MYCN. Theranostics 9:986-1000 (2019). Read more (PubMed: 30867811) »
See all 20 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-4 of 4 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Elena Ezhkova

Verified customer

Submitted Jan 22 2018

Application
ChIP
Detection step
Real-time PCR
Sample
Mouse Cell lysate - nuclear (Min6)
Specification
Min6
Negative control
Negative control for Isl-1 antibody: known locus without enrichment Positive control for IP: Antibody for another factor that is not enriched at that locus
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1.1% Formaldehyed
Positive control
Positive control for Isl-1 antibody: known locus of enrichment Positive control for IP: Antibody for another factor also enriched at that locus

Abcam user community

Verified customer

Submitted Mar 24 2015

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

Abcam user community

Verified customer

Submitted Oct 16 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Sample
Chicken Tissue sections (hindbrain section)
Specification
hindbrain section
Permeabilization
Yes - Triton-X
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 17 2013

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

Sign up