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2 protocols using lamin b1 antibody 12987 1 ap

1

Notch1, EGFR, and β-Catenin Signaling Assays

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Antibodies against Notch1 ICD (D1E11, 1:100 IF, 1:1,000 WB), Notch1 V1744 (D3B8, 1:500 WB), EGFR (D38B1, 1:100 IF, 1:1,000 WB), pEGFR (D7A5, 1:1,000 WB), GFP (D5.1, 1:1,000 WB), non-phospho (active) β-catenin (D13A1, 1:1,000 WB), GAPDH (14C10, 1:10,000 WB), and YAP (D8H1X, 1:200 IF) were from Cell Signaling Technologies. β-catenin antibody (14, 1:1,000 WB) was from BD Biosciences. E-cadherin antibody (HECD-1, 1:1,000 IF, 1:1,000 WB) was from Takara Bio. Notch1 ECD (ABS90, 1:1,000) was from Millipore. FAM83H antibody (1:1,000 WB) was from Bethyl Laboratories. Lamin B1 antibody (12987-1-AP, 1:1,000) was from Proteintech. TexasRed-EGF, rhodamine phalloidin, and Alexa Fluor 488, 568, and 647 goat anti-mouse and anti-rabbit IgG secondary antibodies (1:400) were from Invitrogen. Alexa Fluor 647 azide and EdU cell proliferation kit were from Invitrogen. Hoescht and DAPT were from Sigma. Anti-SNAP (P9310S, 1:1,000), SNAP-Capture, and SNAP-Surface 488 were from New England Biolabs.
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2

Immunofluorescence Analysis of Nuclear Proteins

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The following primary antibodies were used for immunofluorescence experiments: Anti-Huntingtin Protein Antibody, clone mEM48, MAB-5374, Millipore (1:500); lamin B1 Antibody (S-20), sc-30264, Santa Cruz (1:5000); lamin B1 Antibody, 12987-1-AP, Proteintech (1:500); RanGAP1 Antibody (H-180), sc-25630, Santa Cruz (1:500); Anti-GLE1 antibody (ab96007), Abcam (1:200); Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, Millipore (1:500). Secondary antibodies include: Donkey anti Mouse Cy3, Jackson immunology, 715-165-151, (1:500); Goat anti Rabbit Alexa488, Invitrogen, A11008, (1:500); Donkey anti Goat Alexa488, Invitrogen, A11055, (1:500).
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