Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade

ChIP analysis of sheared chromatin from 5x10⁵ HeLaS3 cells using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. Figure 1A. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B. Recovery of the nucleosomes carrying the H3K4me1, H3K4me2, H3K4me3, H3K9me1, H3K27me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K4me1 modification.

ELISA analysis of peptide containing the histone modification of interest using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade.

Dot blot analysis of 0.2 - 100 pmol of the peptides containing other histone modifications and the unmodified H3K4 using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade.
Dilution : 1:5,000

ChIP analysis of sheared chromatin from 1x10⁵ K562 cells using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) 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 3A and B show the H3K4me1 signal in two genomic regions containing the ACTB and GAS2L1 positive controls. The position of the amplicon used for ChIP-qPCR is indicated by an arrow. Figure 3C shows the H3K4me1 peak distribution along a 1 Mb genomic region of chromosome 5.
Antibody amount : 1μg

ChIP analysis of sheared chromatin from 10⁶ K562 cells using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (1 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 gene, respectively, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. Figure 2 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

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 GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade.
Dilution : 1:500

Protein Array analysis of an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade. This figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.
Dilution : 1:2,000

ICC/IF analysis of 4% paraformaldehyde fixed HeLa cells using GTX60818 Histone H3K4me1 (Mono-methyl Lys4) antibody - ChIP grade.
Green : Primary antibody
Blue : DAPI
Dilution : 1:200