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For final validation, we test each antibody using SNAP-ChIP® (Sample Normalization & Antibody Profiling for ChIP) Spike-in Control Panels added to a ChIP experiment. Passing requires that an antibody display <20% cross-reactivity (relative to on-target binding) and >5% enrichment of on-target PTM vs. Input.

Heatmaps are sorted by PTM, and are structured such that the most specific antibodies that passed testing against the full SNAP-ChIP spike-in panel are ranked at the top. These antibodies are recommended for ChIP-seq. The remaining antibodies in the heatmap are separated into three groups: those that failed testing against the full SNAP-ChIP panel, antibodies that were only tested against a subset of spike-ins and passed, and those tested against a subset of spike-ins that failed. Within each subgroup, antibodies are ranked by specificity.

* Denotes antibodies that were tested in Shah et al. 2018.

Efficiency (% Recovered)

< 5% 5-10% > 10%

Efficiency, or enrichment, is the percent recovery of on-target PTM compared to input. Efficiency scores are binned into three categories, shown on the scale. >5% efficiency is required for SNAP-ChIP Certification.

Specificity (% On-Target)

0 20 40 60 80 100 120

Antibody binding is expressed relative to the on-target PTM. Binding is presented as a gradient from blue (weak activity) to red (strong activity). Increasing red across the entire panel denotes antibodies with high cross-reactivity.

K-MetStat

H3K27me1
Vendor: ABclonal Catalog #: A2361 Lot #: 2200190201 Clonality: Polyclonal Species: Rabbit Pass/Fail: Pass # Citations: 2
E0028.3
19.92%
2.17
Unmodified
3.72
H3K4me1
2.72
H3K4me2
3.49
H3K4me3
4.83
H3K9me1
2.67
H3K9me2
2.73
H3K9me3
100.00
H3K27me1
2.54
H3K27me2
2.47
H3K27me3
4.90
H3K36me1
3.58
H3K36me2
4.40
H3K36me3
3.49
H4K20me1
2.24
H4K20me2
2.55
H4K20me3
Vendor: Active Motif Catalog #: 61016 Lot #: 26718014 Clonality: Monoclonal Species: Mouse Pass/Fail: Fail # Citations: 0
E0691.1
18.44%
9.56
Unmodified
6.06
H3K4me1
9.69
H3K4me2
9.17
H3K4me3
21.76
H3K9me1
8.89
H3K9me2
4.47
H3K9me3
100.00
H3K27me1
17.48
H3K27me2
5.38
H3K27me3
4.39
H3K36me1
8.74
H3K36me2
7.23
H3K36me3
3.20
H4K20me1
9.91
H4K20me2
9.95
H4K20me3
Vendor: EpiCypher Catalog #: 13-0015 Lot #: 14247001 Clonality: Polyclonal Species: Rabbit Pass/Fail: Fail # Citations: 0
E0142.1
7.89%
8.09
Unmodified
18.49
H3K4me1
11.51
H3K4me2
13.99
H3K4me3
28.90
H3K9me1
9.80
H3K9me2
32.84
H3K9me3
100.00
H3K27me1
9.56
H3K27me2
7.57
H3K27me3
17.11
H3K36me1
10.85
H3K36me2
16.00
H3K36me3
14.90
H4K20me1
11.89
H4K20me2
10.08
H4K20me3
Vendor: Thermo Fisher Scientific Catalog #: 49-1012 Lot #: A9320234P-190729 Clonality: Polyclonal Species: Rabbit Pass/Fail: Fail # Citations: 1
E0289.1
14.59%
20.86
Unmodified
7.63
H3K4me1
29.70
H3K4me2
22.00
H3K4me3
62.40
H3K9me1
11.57
H3K9me2
6.93
H3K9me3
100.00
H3K27me1
12.87
H3K27me2
6.51
H3K27me3
33.15
H3K36me1
16.56
H3K36me2
14.67
H3K36me3
3.91
H4K20me1
19.77
H4K20me2
18.83
H4K20me3
Vendor: Abclonal Catalog #: A2361 Lot #: 2200019004 Clonality: Polyclonal Species: Rabbit Pass/Fail: Fail # Citations: 0
E0028.2
4.78%
28.21
Unmodified
No Data
H3K4me1
No Data
H3K4me2
No Data
H3K4me3
No Data
H3K9me1
No Data
H3K9me2
No Data
H3K9me3
100.00
H3K27me1
43.33
H3K27me2
29.02
H3K27me3
No Data
H3K36me1
No Data
H3K36me2
No Data
H3K36me3
No Data
H4K20me1
No Data
H4K20me2
No Data
H4K20me3
Vendor: Thermo Fisher Scientific Catalog #: 720097 Lot #: RC226275 Clonality: Polyclonal Species: Rabbit Pass/Fail: Fail # Citations: 0
E0239.1
0.10%
81.73
Unmodified
No Data
H3K4me1
No Data
H3K4me2
No Data
H3K4me3
No Data
H3K9me1
No Data
H3K9me2
No Data
H3K9me3
100.00
H3K27me1
39.33
H3K27me2
51.55
H3K27me3
No Data
H3K36me1
No Data
H3K36me2
No Data
H3K36me3
No Data
H4K20me1
No Data
H4K20me2
No Data
H4K20me3
Efficiency
Unmodified
H3K4me1
H3K4me2
H3K4me3
H3K9me1
H3K9me2
H3K9me3
H3K27me1
H3K27me2
H3K27me3
H3K36me1
H3K36me2
H3K36me3
H4K20me1
H4K20me2
H4K20me3

We use Luminex® multiplexing technology to screen antibody candidates against modified nucleosome substrates. To pass this triage experiment, antibodies must display <10% cross-reactivity relative to the on-target PTM.

Antibodies are sorted in the heatmap by PTM and then by overall specificity, with antibodies that passed Luminex and have highest specificity ranked at the top. Note that passing Luminex does not guarantee an antibody will work well in ChIP experiments; so be sure to confirm that your antibody also passes testing against SNAP-ChIP spike-ins.

Specificity (% On-Target)

0 20 40 60 80 100 120

Antibody binding is expressed relative to the on-target PTM. Binding is presented as a gradient from blue (weak activity) to red (strong activity). Increasing red across the entire panel denotes antibodies with high cross-reactivity.

K-MetStat

H3K27me1
Vendor: ABclonal Catalog #: A2361 Lot #: 2200019004 Clonality: Polyclonal Species: Rabbit Pass/Fail: Pass # Citations: 0
E0028.5
0.10
Unmodified
0.50
H3K4me1
0.30
H3K4me2
0.10
H3K4me3
3.20
H3K9me1
0.10
H3K9me2
0.10
H3K9me3
100.00
H3K27me1
0.40
H3K27me2
0.10
H3K27me3
0.10
H3K36me1
0.20
H3K36me2
0.10
H3K36me3
0.50
H4K20me1
0.10
H4K20me2
0.10
H4K20me3
Vendor: Active Motif Catalog #: 61016 Lot #: 26718014 Clonality: Monoclonal Species: Mouse Pass/Fail: Pass # Citations: 0
E0691.1
0.20
Unmodified
0.20
H3K4me1
0.30
H3K4me2
0.20
H3K4me3
1.40
H3K9me1
0.20
H3K9me2
0.30
H3K9me3
100.00
H3K27me1
3.50
H3K27me2
0.30
H3K27me3
0.30
H3K36me1
0.30
H3K36me2
0.30
H3K36me3
0.20
H4K20me1
0.30
H4K20me2
0.20
H4K20me3
Vendor: Thermo Fisher Scientific Catalog #: 49-1012 Lot #: A9320234P-190729 Clonality: Polyclonal Species: Rabbit Pass/Fail: Pass # Citations: 1
E0289.1
0.50
Unmodified
1.60
H3K4me1
0.70
H3K4me2
0.60
H3K4me3
5.50
H3K9me1
2.00
H3K9me2
0.50
H3K9me3
100.00
H3K27me1
0.80
H3K27me2
0.30
H3K27me3
0.60
H3K36me1
0.60
H3K36me2
1.10
H3K36me3
4.10
H4K20me1
1.00
H4K20me2
1.20
H4K20me3
Vendor: Thermo Fisher Scientific Catalog #: 720097 Lot #: RC226275 Clonality: Polyclonal Species: Rabbit Pass/Fail: Fail # Citations: 0
E0239.1
6.60
Unmodified
8.50
H3K4me1
10.90
H3K4me2
6.10
H3K4me3
6.30
H3K9me1
8.30
H3K9me2
8.20
H3K9me3
100.00
H3K27me1
6.60
H3K27me2
7.90
H3K27me3
5.80
H3K36me1
8.20
H3K36me2
6.70
H3K36me3
7.80
H4K20me1
7.20
H4K20me2
6.70
H4K20me3
Unmodified
H3K4me1
H3K4me2
H3K4me3
H3K9me1
H3K9me2
H3K9me3
H3K27me1
H3K27me2
H3K27me3
H3K36me1
H3K36me2
H3K36me3
H4K20me1
H4K20me2
H4K20me3