ChIP and ChIPseq were performed as described previously (20 (link), 21 (link)). Antibodies used were anti-ASCL1 (Abcam) and anti-IgG (Abcam). ChiPSeq data and full associated methods are both available as an associated file at the GEO database https://www.ncbi.nlm.nih.gov/geo/ and assigned the identifier GSE153823. ChIPseq experiments were performed in two separate clones of each of WT and S-A SH-SY5Y tetracycline-inducible stable cell lines induced with 1 μg/ml doxycycline for 24 hours in four biological replicates. Differential binding analysis (Diffbind) was performed as described previously (20 (link)). ChIP-qPCR primer sequences are provided in Supplementary Table S1 .
Anti ascl1
Manufactured by Abcam
Anti-ASCL1 is a primary antibody that targets the ASCL1 protein, which is a basic helix-loop-helix transcription factor. This antibody can be used in various laboratory techniques to detect and analyze the ASCL1 protein.
Automatically generated - may contain errors
3 protocols using anti ascl1
1
ChIP-Seq Profiling of ASCL1 Binding
2
Immunohistochemistry for Ascl1, Chromogranin, and Synaptophysin
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For immunohistochemistry (IHC), tissue samples were cut as 3 µm thick sections from formalin-fixed paraffin embedded (FFPE) tissues. IHC staining was performed using a Bond Max automated staining system (Leica) using the antibodies: anti-Ascl1 (Abcam #211327, 1:400), anti-Chromogranin (Abcam #52983, 1:250), anti-Synaptophysin (Abcam #32127, 1:1000). GLuc and Cas9 staining was performed manually using the antibodies: anti-GLuc (Prolume Ltd, 1:1000), anti-Cas9 (Cell Signaling #19526, 1:400). Images were acquired using the Leica Aperio Versa slide-scanner and Leica Aperio eSlide Manager software v. 1.0.3.37. IHC images were analyzed quantitatively using the Aperio ImageScope software v. 12.3.2.8013. Tumors were marked and outlined by individuals blinded to the experimental setup and their area quantified in ImageScope. Tumor burden was calculated as percentage of tumor area to total lung area.
3
Immunohistochemistry for Neural Markers
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