Anti-Islet 1 antibody - Neural Stem Cell Marker (ab20670)

Overview

  • Product nameAnti-Islet 1 antibody - Neural Stem Cell Marker
    See all Islet 1 primary antibodies
  • Description
    Rabbit polyclonal to Islet 1 - Neural Stem Cell Marker
  • Specificityab20670 might also detect rat and human Islet 2 protein as the immunogen used to raise this ab20670 is 86% identical to rat and human Islet 2. This has not been tested.
  • Tested applicationsSuitable for: IHC-FoFr, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Apteronotus leptorhynchus
    Predicted to work with: Chicken, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Islet 1.

    (Peptide available as ab21996.)

  • Positive control
    • IHC-P: FFPE mouse embryo E12.

Properties

Applications

Our Abpromise guarantee covers the use of ab20670 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-FoFr Use at an assay dependent concentration. PubMed: 20096094
IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 2 µg/ml.
IHC-Fr 1/500.

Target

Anti-Islet 1 antibody - Neural Stem Cell Marker images

  • IHC image of Islet 1 staining in a section of formalin fixed, paraffin embedded mouse embryo E12, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval (EDTA based pH 9.0 solution, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab20670, 0.5µg/ml, 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.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • IHC image of Islet 1 staining in a section of formalin fixed, paraffin embedded mouse embryo E12, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval (EDTA based pH 9.0 solution, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab20670, 0.5µg/ml, 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.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab20670 2µg/ml staining ISLET1 in human pancreas using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and weak cytoplasmic staining primarily in the pancreatic islet.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Dorsal root ganglion explants were dissected from 16 day-old rat embryos and cultured for 6 hours in vitro with Neurobasal Medium containing B27 supplement.     

    Nuclei stained positive for anti-Islet 1 antibody ab20670 at 2µg/ml. As would be expected, not all cells in this preparation were Islet 1-positive. Pre-incubation of ab20670 with the immunizing peptide ab21996 resulted in complete blocking of the antibody.

    Green = ab20670
    Blue = To-pro-3 nuclear stain

    The level of magnification is different in each image.    

  • Immunocytochemistry/ Immunofluorescence analysis of adult mouse cardiac fibroblasts labeling Islet 1 with ab20670 at 1/100 dilution (green). Samples were fixed using 4% PFA with 0.25% TritonX-100. Scale bar is 100 µM.

  • Immunohistochemistry (Frozen sections) analysis of mouse brain tissue sections labeling Islet 1 with ab20670 at 1/200 dilution. The tissue was fixed with paraformaldehyde followed by blocking with 5% serum for 1 hour at 25°C. The tissue was incubated with ab20670 in PBST for 12 hours at 4°C. A polyclonal donkey anti-rabbit Alexa Fluor® 594 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of mouse dorsal root ganglia (DRG) neurons labeling Islet 1 with ab20670 at 1/200 dilution. Cells were fixed with paraformaldehyde and permeabilized with Tween 20. Blocking of the cells was performed with 2% serum for 30 minutes at 21°C, followed by incubation with ab20670 for 18 hours at 4°C. A polyclonal goat anti-rabbit Alexa Fluor® 568 secondary antibody was used at 1/1000 dilution. DAPI was used to counterstain.

    See Abreview

  • Immunohistochemical analysis of paraffin-embedded mouse E11.5 somites labelling Islet 1 with ab20670 at 1/100 dilution, followed by BioGenex polymer-HRP reagent (undiluted according to manufacturer).

  • ab20670 detected Islet 1 in the nucleus of frozen sections of E11.5 mouse embryonic brain using 1 ug/ml of antibody. Brighter staining may be achieved using a greater concentration of antibody, e.g. 2.5 or 5 ug/ml.

References for Anti-Islet 1 antibody - Neural Stem Cell Marker (ab20670)

This product has been referenced in:
  • Nishi K  et al. Nardilysin Is Required for Maintaining Pancreatic ß-Cell Function. Diabetes 65:3015-27 (2016). Read more (PubMed: 27385158) »
  • Synnergren J  et al. Comparative transcriptomic analysis identifies genes differentially expressed in human epicardial progenitors and hiPSC-derived cardiac progenitors. Physiol Genomics 48:771-784 (2016). ICC/IF ; Human . Read more (PubMed: 27591124) »

See all 23 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Mouse Cell (Differentiated mES cell)
Permeabilization Yes - 0.2% Triton X
Specification Differentiated mES cell
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 23 2016

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

Abcam user community

Verified customer

Submitted Jan 12 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Heat mediated
Sample Spotted Catshark (Scyliorhinus canicula) Tissue sections (brain cells)
Specification brain cells
Permeabilization No
Fixative Paraformaldehyde
Username

Dr. Liam Rasch

Verified customer

Submitted Nov 05 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Dorsal Root Ganglia (DRG) neurons)
Specification Dorsal Root Ganglia (DRG) neurons
Fixative Paraformaldehyde
Permeabilization Yes - tween 20
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 21°C
Username

Abcam user community

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Submitted Oct 18 2012

Application Western blot
Sample Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount 50 µg
Specification Brain
Gel Running Conditions Reduced Denaturing (4-15%)
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Sep 28 2012

Thank you for your message.

I am pleased the results have improved. In order to help further, I would appreciate if you are able to send an attached image which will help me to assess the results.

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Please find attached the Certificate of Compliance for ab20670, lot #*.

If there is anything else I can help you with, please let me know.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Adult heart)
Specification Adult heart
Fixative Formaldehyde
Permeabilization Yes - 0.5% Tx100 in PBS
Blocking step Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1% · Temperature: 20°C
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Submitted Jun 19 2012

1-10 of 22 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"