Anti-Islet 1 antibody - Neural Stem Cell Marker

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

• IHC-Fr

AbReview23021****

Immunohistochemistry (Frozen sections) - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

This image is courtesy of an anonymous abreview.

• ICC/IF

AbReview32444****

Immunocytochemistry/ Immunofluorescence - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

This image is courtesy of an anonymous abreview.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

Image from Ifkovits, Jamie L. et al. PLoS ONE 9.2 (2014): e89678. doi: 10.1371/journal.pone.0089678. Fig S3B. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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.

This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

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).

This image is courtesy of an anonymous collaborator.

• IHC

CiteAb

Immunohistochemistry - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

Immunohistochemistry using Anti-Islet 1 antibody, ab20670. Publication image from Muzio, L. et al., 2020, Nat Commun, 32737286. Legend direct from paper.

VPS35 levels in ALS iPSCs-derived MNs and postmortem biopsies.a, b IHC for VPS35 in alpha-MNs in representative sections from ventral horns of SCs biopsies from a non-neurological control (a) and an ALS patient (b). c, d, IHC for VPS26 in alpha-MNs sampled in adjacent sections from a non-neurological control (c) and an ALS patient (d). Analyses were done on four non-neurological control biopsies and five ALS biopsies examined over three independent experiments. e Maximum projections of representative confocal stacks (three independent experiments) for VPS35 and ISLET1 in representative cultures of iPSC-derived MNs from a healthy volunteer (#8) and an ALS patient (#13SOD1Leu144Phe). A representative WB for VPS35 and β-Actin in protein extracts from iPSCs-derived MN cultures (#8 SOD1Asn65Ser, #13SOD1Leu144Phe, #27 SOD1Asp97Asn and age- and sex-matched healthy volunteers #2, #4, and #8) is shown in f. Quantifications derived from n = 3 controls (#2, #4, and #8) and from n = 3 ALS (#8, #13, and #27), means (±SD) are examined from two independent experiments (p = 0.005). g Representative cross-sectional analysis of confocal stacks (three independent experiments) for VPS35 and ISLET1 in cultured iPSCs-derived MNs from an healthy volunteer (#8) and from an ALS patient (#27 SOD1Asp97Asn) with or without compound 2a (10 µM for 6 days). h Representative WB for VPS35 and β-Actin in iPSCs-derived MNs of an ALS patient (#27) treated with vehicle or with compound 2a (10 µM for 6 days). Quantifications (means ± SD) derived from n = 3 wells/treatment and data are examined from one experiment (p = 0.017). Two-tailed Student’s t-test was used to determine the statistical significance in f and h. **p < 0.01. Scale bar : 150 µm in d, 25 µm in e, 15 µm in g.

• IHC

CiteAb

Immunohistochemistry - Anti-Islet 1 antibody - Neural Stem Cell Marker (AB20670)

Immunohistochemistry using Anti-Islet 1 antibody, ab20670. Publication image from Muzio, L. et al., 2020, Nat Commun, 32737286. Legend direct from paper.

VPS35 levels in ALS iPSCs-derived MNs and postmortem biopsies.a, b IHC for VPS35 in alpha-MNs in representative sections from ventral horns of SCs biopsies from a non-neurological control (a) and an ALS patient (b). c, d, IHC for VPS26 in alpha-MNs sampled in adjacent sections from a non-neurological control (c) and an ALS patient (d). Analyses were done on four non-neurological control biopsies and five ALS biopsies examined over three independent experiments. e Maximum projections of representative confocal stacks (three independent experiments) for VPS35 and ISLET1 in representative cultures of iPSC-derived MNs from a healthy volunteer (#8) and an ALS patient (#13SOD1Leu144Phe). A representative WB for VPS35 and β-Actin in protein extracts from iPSCs-derived MN cultures (#8 SOD1Asn65Ser, #13SOD1Leu144Phe, #27 SOD1Asp97Asn and age- and sex-matched healthy volunteers #2, #4, and #8) is shown in f. Quantifications derived from n = 3 controls (#2, #4, and #8) and from n = 3 ALS (#8, #13, and #27), means (±SD) are examined from two independent experiments (p = 0.005). g Representative cross-sectional analysis of confocal stacks (three independent experiments) for VPS35 and ISLET1 in cultured iPSCs-derived MNs from an healthy volunteer (#8) and from an ALS patient (#27 SOD1Asp97Asn) with or without compound 2a (10 µM for 6 days). h Representative WB for VPS35 and β-Actin in iPSCs-derived MNs of an ALS patient (#27) treated with vehicle or with compound 2a (10 µM for 6 days). Quantifications (means ± SD) derived from n = 3 wells/treatment and data are examined from one experiment (p = 0.017). Two-tailed Student’s t-test was used to determine the statistical significance in f and h. **p < 0.01. Scale bar : 150 µm in d, 25 µm in e, 15 µm in g.