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Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Islet 1.
(Peptide available as ab21996.)
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.
|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 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.
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.
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.
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).