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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 orange (weak activity) to dark blue (strong activity). Increasing blue across the entire panel denotes antibodies with high cross-reactivity.

OncoStat

H3.3K27M
Vendor: RevMAb Catalog #: 31-1175-00 Lot #: S-07-02376 Clonality: Monoclonal Species: Rabbit Pass/Fail: Pass # Citations: 2
E0625.1
73.27%
0.14
H3.3
0.05
H3.3K4M
0.09
H3.3K9M
100.00
H3.3K27M
0.15
H3.3G34R
0.30
H3.3G34V
0.11
H3.3G34W
0.22
H3.3K36M
Vendor: Thermo Fisher Scientific Catalog #: MA5-27916 Lot #: UG2809141 Clonality: Monoclonal Species: Rabbit Pass/Fail: Pass # Citations: 0
E0771.3
73.27%
0.14
H3.3
0.05
H3.3K4M
0.09
H3.3K9M
100.00
H3.3K27M
0.15
H3.3G34R
0.30
H3.3G34V
0.11
H3.3G34W
0.22
H3.3K36M
Vendor: Thermo Fisher Scientific Catalog #: MA5-27916 Lot #: UI2836151 Clonality: Monoclonal Species: Rabbit Pass/Fail: Pass # Citations: 0
E0771.4
73.27%
0.14
H3.3
0.05
H3.3K4M
0.09
H3.3K9M
100.00
H3.3K27M
0.15
H3.3G34R
0.30
H3.3G34V
0.11
H3.3G34W
0.22
H3.3K36M
Vendor: Thermo Fisher Scientific Catalog #: MA5-27916 Lot #: UI2840791 Clonality: Monoclonal Species: Rabbit Pass/Fail: Pass # Citations: 0
E0771.5
73.27%
0.14
H3.3
0.05
H3.3K4M
0.09
H3.3K9M
100.00
H3.3K27M
0.15
H3.3G34R
0.30
H3.3G34V
0.11
H3.3G34W
0.22
H3.3K36M
Vendor: EMD Millipore Catalog #: ABE419 Lot #: 2738997 Clonality: Polyclonal Species: Rabbit Pass/Fail: Fail # Citations: 44
E0272.1
1.18%
0.68
H3.3
0.31
H3.3K4M
0.28
H3.3K9M
100.00
H3.3K27M
1.52
H3.3G34R
0.58
H3.3G34V
0.74
H3.3G34W
0.59
H3.3K36M
Efficiency
H3.3
H3.3K4M
H3.3K9M
H3.3K27M
H3.3G34R
H3.3G34V
H3.3G34W
H3.3K36M

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 orange (weak activity) to dark blue (strong activity). Increasing blue across the entire panel denotes antibodies with high cross-reactivity.

There are no tests for this method.