Journal:

Article Title: Modular Structure of the Human Lamin B2 Replicator

doi: 10.1128/MCB.24.7.2958-2967.2004

Figure Lengend Snippet: ChIP analysis. H30 cells containing eLamB2* were cross-linked with formaldehyde, and chromatin was immunoprecipitated with an antibody against hOrc2 (α-ORC2), with a control antibody against transcription factor USF-1 (α-USF-1) and with a control nonspecific antibody (IgG). In 24.79 cells containing eLamB2-mut, ChIP was performed with α-ORC2 only. (A) Immunoprecipitation was verified by Western blot analysis. Input, chromatin before immunoprecipitation; IP, immunoprecipitated material. Samples were probed with the antibody indicated on the right of each panel. The star identifies the heavy chain of the antibody. (B) Quantification of DNA immunoprecipitated from H30 and 24.79 cells. After reversion of cross-linking, immunoprecipitated DNA was subjected to competitive PCR with primers B48 and B13. Competitor dilutions are indicated above each lane. The identity of the different amplification bands is indicated on the right of each panel. (C) In the case of the H30 clone, histograms show the enrichment of endogenous (Ori) and ectopic (e-Ori*) sequences over B13 in the pool of DNA immunopurified with α-ORC2, α-USF-1, or control (IgG) antibodies. For clone 24.79, the histogram shows the enrichment of ectopic (e-Ori-mut) over endogenous B13 sequences immunopurified with α-ORC2 antibody. Bars indicate variations in three independent experiments.

Article Snippet: Immunoprecipitations were performed by incubation with 10 μg of anti-USF-1 (Santa Cruz Biotechnology) rabbit polyclonal antibody H-86, 10 μg of anti-ORC2 (StressGen Biotechnology) monoclonal antibody, and 10 μg of anti-immunoglobulin G (IgG) (Santa Cruz Biotechnology) antibody as negative control.

Techniques: Immunoprecipitation, Western Blot, Amplification