Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody

Western Blot: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - Western blot was performed on whole cell extracts (25 ug, lane 1) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the antibody against H2A.Zac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left.

Immunocytochemistry/Immunofluorescence: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - HeLa cells were stained with the antibody against H2A.Zac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2AZac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Chromatin Immunoprecipitation: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - ChIP assays were performed using human K562 cells, the antibody against H2AZac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes CCT5 and EIF4A2, used as positive controls, and for the coding region of the inactive MYT1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a percent of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis