Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody - BSA Free

Western Blot: Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252]

Western Blot: Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H4K5,8,12ac . 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 (kDa) is shown on the left.

Immunocytochemistry/Immunofluorescence: Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - HeLa cells were stained with Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody and DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. (Top): cells were immunofluorescently labeled with the Histone H4 [ac Lys5, ac Lys8, ac Lys12] 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. Subsequent images show staining of the cells with the Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody after incubation of the antibody with 10 ng/ul of the following blocking peptides: H4K5,8,12 unmodified, H4K5,8,12ac, H2A.ZK5,7,11ac and H4K5,8,12,16ac, respectively.

Chromatin Immunoprecipitation: Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - ChIP assays were performed using human K562 cells, the antibody against H4K5,8,12ac and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP- seq kit on 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. QPCR was performed with primers for promoter of the active gene EIF4A2 and for a region 1 kb upstream of the GAPDH gene, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).