WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using GTX60335 Histone H4K5ac (acetyl Lys5) antibody - ChIP grade.
Dilution : 1:500

Dot blot analysis of 0.2 - 100 pmol of the peptides containing other histone modifications and the unmodified H4 using GTX60335 Histone H4K5ac (acetyl Lys5) antibody - ChIP grade.
Dilution : 1:5,000

ICC/IF analysis of 4% paraformaldehyde fixed HeLa cells using GTX60335 Histone H4K5ac (acetyl Lys5) antibody - ChIP grade.
Green : Primary antibody
Blue : DAPI
Dilution : 1:500

ChIP analysis of sheared chromatin from 10⁶ HeLa cells using GTX60335 Histone H4K5ac (acetyl Lys5) antibody - ChIP grade. A titration of the antibody consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (2 μg/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. This figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ChIP analysis of sheared chromatin from 10⁶ HeLa cells using GTX60335 Histone H4K5ac (acetyl Lys5) antibody - ChIP grade. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the signal distribution along the complete length of chromosome 1 (figure 2A) and a zoomin to a 500 kb region (figure 2B). Figure 2C and D show the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls. The position of the amplicon used for validating the QPCR results is shown with an arrow
Antibody amount : 1μg

ELISA analysis of peptide containing the histone modification of interest using GTX60335 Histone H4K5ac (acetyl Lys5) antibody - ChIP grade.