Antibody Validation Database

An assessment of histone-modification antibody quality

Description

We have tested the specificity and utility of more than 200 antibodies raised against 57 different histone modifications in Drosophila melanogaster, Caenorhabditis elegans and human cells. Although most antibodies performed well, more than 25% failed specificity tests by dot blot or western blot. Among specific antibodies, more than 20% failed in chromatin immunoprecipitation experiments. We advise rigorous testing of histone-modification antibodies before use and provide a website for posting new test results (http://compbio.med.harvard.edu/antibodies/) [Published in Nature Structural & Molecular Biology, 2010]

Targets (62)

Name	Target Species	Antibodies	Validated Species	Validations? W: Western blot validations D: Dot blot validations C: ChIP-chip/seq validataions	Passed / Total? Western blot: Pass if the histone band constituted at least 50% of the total nuclear signal, was at least ten-fold more intense than any other single nuclear band, and was at least ten-fold more intense than recombinant, unmodified histone. Dot blot: Pass if at least 75% of the total signal was specific to the cognate peptide. ChIP-chip/seq: Pass if the Pearson correlation is more than 0.8 on any pair of ChIPs performed from independent preparations matched for stage, cell type or biological tissue
CitH3	multiple	1	fruit fly	W100	0 / 1
H2AK5ac	multiple	3	fruit fly, human	W3D2C2	4 / 7
H2AK7ac	multiple	1	fruit fly	W100	0 / 1
H2AS129ph	multiple	1	human	W1D1C1	1 / 3
H2B	multiple	1	fruit fly	W10C1	1 / 2
H2BK120ac	multiple	1	human	W1D1C1	2 / 3
H2BK15ac	multiple	1	human	W1D1C1	2 / 3
H2BK16ac	multiple	1	fruit fly	W100	0 / 1
H2BK5ac	multiple	4	fruit fly, human	W4D3C2	3 / 9
H2BK5me1	multiple	1	human	W1D1C1	1 / 3
H2Bac	multiple	1	fruit fly	W100	0 / 1
H3	multiple	4	fruit fly, worm	W40C3	6 / 7
H3K14ac	multiple	3	fruit fly, human	W3D1C3	4 / 7
H3K14acS10ph	multiple	1	fruit fly	W10C1	1 / 2
H3K18ac	multiple	4	fruit fly, human, multiple	W4D4C4	11 / 12
H3K23ac	multiple	1	fruit fly, human	W2D1C2	5 / 5
H3K23me2	multiple	1	human	W1D1C1	2 / 3
H3K27ac	multiple	9	fruit fly, human, worm	W9D6C6	14 / 21
H3K27me1	multiple	8	human, multiple, worm	W7D8C5	11 / 20
H3K27me2	multiple	9	fruit fly, human, multiple, worm	W6D6C4	5 / 16
H3K27me3	multiple	11	fruit fly, human, multiple, worm	W10D7C8	15 / 25
H3K36ac	multiple	2	multiple, worm	W1D3C1	3 / 5
H3K36me1	multiple	6	fruit fly, human, multiple, worm	W6D6C3	9 / 15
H3K36me2	multiple	6	fruit fly, human, worm	W6D5C4	7 / 15
H3K36me3	multiple	8	fruit fly, human, worm	W7D7C8	17 / 22
H3K4ac	multiple	1	human	W1D1C1	2 / 3
H3K4me1	multiple	8	fruit fly, human, multiple, worm	W8D7C8	17 / 23
H3K4me2	multiple	8	fruit fly, human, worm	W8D7C6	17 / 21
H3K4me3	multiple	10	fruit fly, human, worm	W8D8C9	22 / 25
H3K56ac	multiple	3	fruit fly, human	W3D2C2	4 / 7
H3K79me1	multiple	4	fruit fly, human, worm	W5D4C5	9 / 14
H3K79me2	multiple	5	fruit fly, human, worm	W4D4C4	10 / 12
H3K79me3	multiple	4	fruit fly, human, multiple, worm	W3D4C2	4 / 9
H3K9ac	multiple	7	fruit fly, human, multiple, worm	W7D4C5	10 / 16
H3K9acS10ph	multiple	6	fruit fly, multiple	W5D4C2	9 / 11
H3K9me1	multiple	9	fruit fly, human, worm	W9D6C7	15 / 22
H3K9me2	multiple	11	fruit fly, human, worm	W9D8C8	14 / 25
H3K9me3	multiple	16	fruit fly, human, multiple, worm	W15D10C10	25 / 35
H3R17me2a	multiple	5	fruit fly, human	W6D3C1	1 / 10
H3R26me2a	multiple	1	human	W1D10	0 / 2
H3R2me1	multiple	1	fruit fly, human	W2D10	0 / 3
H3R2me2a	multiple	3	human	W3D30	3 / 6
H3R2me2s	multiple	1	human	W1D10	1 / 2
H3R8me1	multiple	1	human	W1D1C1	1 / 3
H3R8me2a	multiple	1		0D10	0 / 1
H3S10ph	multiple	8	fruit fly, human, worm	W8D4C2	7 / 14
H3S28ph	multiple	5	fruit fly, worm	W50C2	3 / 7
H3T11ph	multiple	2	human	W2D2C1	3 / 5
H4	multiple	3	fruit fly, worm	W20C1	3 / 3
H4K12ac	multiple	3	fruit fly, human	W3D2C3	5 / 8
H4K16ac	multiple	8	fruit fly, multiple, worm	W5D5C3	8 / 13
H4K20me1	multiple	9	fruit fly, human, multiple, worm	W7D8C4	17 / 19
H4K20me2	multiple	3	fruit fly, worm	W2D10	2 / 3
H4K20me3	multiple	11	fruit fly, human, multiple, worm	W11D7C4	3 / 22
H4K5ac	multiple	2	fruit fly, human	W3D2C3	7 / 8
H4K8ac	multiple	4	fruit fly, human, worm	W2D4C3	8 / 9
H4K91ac	multiple	2	fruit fly, human	W2D1C1	2 / 4
H4R3me1	multiple	1	human	W1D10	1 / 2
H4R3me2	multiple	2	fruit fly	W200	0 / 2
H4R3me2a	multiple	1	human	W1D10	0 / 2
H4R3me2s	multiple	1	human	W1D10	1 / 2
H4ac4	multiple	2	fruit fly, worm	W10C2	3 / 3