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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

H3K4me0
Vendor: EMD Millipore Catalog #: 05-1341 Lot #: 2817186 Clonality: Monoclonal Species: Mouse Pass/Fail: Fail # Citations: 13
E0626.1*
5.03%
100.00
Unmodified
31.74
H3K4me1
32.66
H3K4me2
21.30
H3K4me3
No Data
H3K9me1
No Data
H3K9me2
No Data
H3K9me3
No Data
H3K27me1
No Data
H3K27me2
No Data
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

H3K4me0
Vendor: RevMAb Catalog #: 31-1086-00 Lot #: Q-03-00944 Clonality: Monoclonal Species: Rabbit Pass/Fail: Fail # Citations: 0
E0613.1
100.00
Unmodified
100.00
H3K4me1
88.90
H3K4me2
87.50
H3K4me3
61.10
H3K9me1
106.30
H3K9me2
97.20
H3K9me3
109.00
H3K27me1
94.40
H3K27me2
75.70
H3K27me3
111.10
H3K36me1
86.10
H3K36me2
88.20
H3K36me3
84.70
H4K20me1
102.80
H4K20me2
97.90
H4K20me3
Unmodified
H3K4me1
H3K4me2
H3K4me3
H3K9me1
H3K9me2
H3K9me3
H3K27me1
H3K27me2
H3K27me3
H3K36me1
H3K36me2
H3K36me3
H4K20me1
H4K20me2
H4K20me3