Journal: PLoS ONE

Article Title: Loss of the Histone Pre-mRNA Processing Factor Stem-Loop Binding Protein in Drosophila Causes Genomic Instability and Impaired Cellular Proliferation

doi: 10.1371/journal.pone.0008168

Figure Lengend Snippet: Protein lysates from wt, Slbp 15 , Slbp 10 , and H2aV 810 mutants were obtained from whole 3 rd instar larvae and probed with antibodies to A) H2b and H3, and B) H3K4-me2 and H3K9-me2. β-tubulin was used as a loading control for both panels.

Article Snippet: Polyclonal rabbit C-terminal H3 (1∶3000) (Abcam #1791), polyclonal rabbit H2b (Abcam #1790) (1∶3000), monoclonal mouse H3K9-me2 (Abcam #1220) (1∶750), and polyclonal rabbit H3K4-me2 (Abcam #7766) (1∶3000), and monoclonal mouse Β-tubulin (1∶1000) antibodies were obtained commercially.

Techniques:

Journal: bioRxiv

Article Title: Fundamental Role Of The H2A.Z C-Terminal Tail In The Formation Of Constitutive Heterochromatin

doi: 10.1101/2021.02.22.432230

Figure Lengend Snippet: Intranuclear heterogeneity of H2A.Z-containing nucleosomes. (A) Comparison of the salt elution profiles of H2A, H2A.X, H2A.Z detected by ZAbA (Abcam, ab 97966) antibody and of H3-GFP (used as an internal control) in HeLa cell nuclei. (B) Salt elution curves of H2A.Z detected by H2A.Z specific polyclonal antibodies from different manufacturers: ZAbA, ZAbB (Thermo Fisher Sci., PA5-17336). (C) CLSM images and line-scans showing nuclear localization of H2A.Z recognized by the ZAbA antibody (upper panel), or by ZAbB (lower panel). (D) Flow chart of the immuno-cross-linking (immune-X-linking) experiment in panels E, F. (E and F) immune-X-linking of H2A.Z immobilizes the H3K9me3 containing nucleosomes (E), in contrast with the H3K27me3 containing nucleosomes (F), measured in H2A.Z1ΔC (ΔC) and H2A.Z1 (control; CTRL) expressing DKO DT40 cells.

Article Snippet: The blocking solution was washed out with 500 μl ice cold 1×PBS/EDTA 3 times for 3 minutes and indirect immunofluorescence labeling was performed using rabbit polyclonal anti-H2A.Z (ab97966 (ZAbA), ab4174, Abcam, Cambridge, UK; 1 mg/ml), sheep polyclonal anti-H2A.Z (acetyl K4+K7+K11, ab18262, Abcam, Cambridge, UK; 0.5 mg/ml), rabbit polyclonal anti-H2A.Z (PA5-17336 (ZAbB), Thermo Fisher Scientific, Waltham, Massachusetts, USA), rabbit polyclonal anti-PWWP2A (NBP2-13833, Novus Biologicals, Centennial, Colorado, USA; 0.2 mg/ml), rabbit polyclonal anti-H2A.X (ab11175, Abcam, Cambridge, UK; 1 mg/ml), rabbit polyclonal anti-H2A (ab18255, Abcam, Cambridge, UK; 1 mg/ml), rabbit polyclonal anti-H2B (ab52484, Abcam, Cambridge, UK; 1 mg/ml), mouse monoclonal anti-H1 (ab71591, Abcam, Cambridge, UK; 1 mg/ml), mouse monoclonal anti-HP1 (ab234085, Abcam, Cambridge, UK; 1 mg/ml), mouse monoclonal anti-γH2A.X (05-636, Merck-Millipore, Darmstadt, Germany), mouse monoclonal anti-H3K4me3 (( ); 0,5 mg/ml), mouse monoclonal anti-H3K9me3 (( ); 0,5 mg/ml) or mouse monoclonal anti-H3K27me3 (( ); 0,5 mg/ml) primary antibodies, all diluted in 150 μl of 1×PBS/EDTA/1% BSA (1×PBS/EDTA supplemented with 1% w/v bovine serum albumin), at 4°C, overnight.

Techniques: Expressing