ICC/IF analysis of paraformaldehyde fixed HeLa asynchronous cells using GTX60329 Histone H3K27me3S28ph (Tri-methyl Lys27/phospho Ser28) antibody - ChIP grade. Phosphorylation of H3 on serine 28 is increased during late G2 phase and reaches a maximum in metaphase cells. This may explain the different staining intensities of different cells.
Green : Primary antibody
Blue : DAPI
Dilution : 1:200

ChIP analysis of sheared chromatin from 10⁴ HeLa cells treated with colcemid using GTX60329 Histone H3K27me3S28ph (Tri-methyl Lys27/phospho Ser28) antibody - ChIP grade. A titration of the antibody consisting of 1, 5 and 10 μl per ChIP experiment was analysed. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active genes c-fos and RPL30 and for the inactive gene MYOD. This figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Dot blot analysis of 0.2 - 100 pmol of the peptides containing other modifications of histone H3 and H4 and unmodified sequences from histone H3 using GTX60329 Histone H3K27me3S28ph (Tri-methyl Lys27/phospho Ser28) antibody - ChIP grade.
Dilution : 1:20,000

WB analysis of histone extracts (15 μg) from HeLa cells using GTX60329 Histone H3K27me3S28ph (Tri-methyl Lys27/phospho Ser28) antibody - ChIP grade.
Dilution : 1:250

IP analysis of HeLa cells were treated with colcemid (to block the cell cycle in metaphase) using GTX60329 Histone H3K27me3S28ph (Tri-methyl Lys27/phospho Ser28) antibody - ChIP grade.
IP reaction : 5 μl antibody / 1x10⁴ cells
Lane 1 : Primary antibody
Lane 2 : No antibody control
Lane 3 : Input
Dilution : 1:500

ELISA analysis of peptide containing the histone modifications of interest using GTX60329 Histone H3K27me3S28ph (Tri-methyl Lys27/phospho Ser28) antibody - ChIP grade.