macro-H2A1.2 antibody | knockout validation | Millipore MABE61

This is a knockout-validated antibody summary, based on the publication "Replication Stress Shapes a Protective Chromatin Environment across Fragile Genomic Regions", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Mouse monoclonal IgG2bλ

Company: Millipore

Antibody: macro-H2A1.2

Catalog number: MABE61

Summary: Mouse monoclonal IgG2bλ against a recombinant protein corresponding to human macro-H2A1.2. Reacts with human by western blot, immunocytochemistry and immunohistochemistry.

Validation Method

Western blot

Sample

WT and macroH2A1.2 knockout (KO) K562 cells. Cells were resuspended in 0.5 mL lysis buffer containing 10 mM HEPES (pH 7.4), 10 mM KCl, 0.5% NP-40 with phosphatase inhibitors (Calbiochem) and after separation of the nucleoplasmic fraction, chromatin-bound proteins were released from the DNA by addition of 0.2 N HCl.

Secondary incubation

Goat anti-mouse IgG-HRP (Santa Cruz sc-2005, Invitrogen 31430).

Detection

Enhanced chemiluminescence (Amersham).

Figure

Western blot analysis of chromatin-bound (Chr) and non-chromatin-bound (unbound) protein fractions from K562 WT or macroH2A1.2 knockout (KO) cells after the indicated treatments. Please see Figure 3a in the article [1].

References
  1. Kim J, Sturgill D, Sebastian R, Khurana S, Tran A, Edwards G, et al. Replication Stress Shapes a Protective Chromatin Environment across Fragile Genomic Regions.. Mol Cell. 2018;69:36-47.e7 pubmed publisher