List of Targets (121 total)
Target | HGNC Symbol | Synonym | Assay Format | Reader domains present in construct |
---|---|---|---|---|
ASH1L-[BRD]-GST | ASH1L | Bromodomain-containing ASH-1 like protein | ThermoShift | BRD |
ATAD2-His | ATAD2 | ATPase family AAA domain-containing protein 2 | ThermoShift; AlphaScreen | BRD |
ATAD2B-GST | ATAD2B | ATPase family AAA domain-containing protein 2B | ThermoShift | BRD |
ATAD2B-His | ATAD2B | ATPase family AAA domain-containing protein 2B | ThermoShift; AlphaScreen | BRD |
BAZ1A-GST | BAZ1A | hACF1; WCRF180; hWALp1 | ThermoShift | BRD |
BAZ1B-His | BAZ1B | Bromodomain adjacent to zinc finger domain protein 1B | ThermoShift; AlphaScreen | BRD |
BAZ2A-GST | BAZ2A | Bromodomain adjacent to zinc finger domain protein 2A | ThermoShift | BRD |
BAZ2A-His | BAZ2A | Bromodomain adjacent to zinc finger domain protein 2A | AlphaScreen | BRD |
BAZ2B-His | BAZ2B | Bromodomain adjacent to zinc finger domain protein 2B | ThermoShift; AlphaScreen | BRD |
BPTF-[BRD]-His | BPTF | Bromodomain-containing transcription factor | ThermoShift; AlphaScreen | BRD |
BPTF-[PHD-BRD]-His | BPTF | Bromodomain and PHD finger-containing transcription factor | AlphaScreen; ThermoShift | BRD; PHD |
BRD1-GST | BRD1 | Bromodomain-containing protein 1 | ThermoShift | BRD |
BRD1-His | BRD1 | Bromodomain-containing protein 1 | ThermoShift; AlphaScreen | BRD |
BRD2-1-GST | BRD2 | Bromodomain-containing protein 2 domain 1 | ThermoShift | BRD |
BRD2-1-His | BRD2 | Bromodomain-containing protein 2 domain 1 | ThermoShift; AlphaScreen | BRD |
BRD2-2-GST | BRD2 | Bromodomain-containing protein 2 domain 1 | ThermoShift | BRD |
BRD2-2-His | BRD2 | Bromodomain-containing protein 2 domain 2 | ThermoShift; AlphaScreen | BRD |
BRD2-Tndm-His | BRD2 | Bromodomain-containing protein 2 domain 1 and domain 2 | ThermoShift; AlphaScreen | BRD |
BRD3-1-GST | BRD3 | Bromodomain-containing protein 3 domain 1 | ThermoShift | BRD |
BRD3-1-His | BRD3 | Bromodomain-containing protein 3 domain 1 | ThermoShift; AlphaScreen | BRD |
BRD3-2-GST | BRD3 | Bromodomain-containing protein 3 domain 2 | ThermoShift | BRD |
BRD3-2-His | BRD3 | Bromodomain-containing protein 3 domain 2 | ThermoShift; AlphaScreen | BRD |
BRD3-Tndm-GST | BRD3 | Bromodomain-containing protein 3 domain 1 and domain 2 | ThermoShift | BRD |
BRD3-Tndm-His | BRD3 | Bromodomain-containing protein 3 domain 1 and domain 2 | ThermoShift; AlphaScreen | BRD |
BRD4 Full length | BRD4 | Bromodomain-containing protein 4, full length | ThermoShift AlphaScreen | BRD |
BRD4-1 | BRD4 | Bromodomain-containing protein 4 domain 1 | SPR | BRD |
BRD4-1-GST | BRD4 | Bromodomain-containing protein 4 domain 1 | ThermoShift | BRD |
BRD4-1-His | BRD4 | Bromodomain-containing protein 4 domain 1 | ThermoShift; AlphaScreen | BRD |
BRD4-2 | BRD4 | Bromodomain-containing protein 4 domain 2 | SPR | BRD |
BRD4-2-GST | BRD4 | Bromodomain-containing protein 4 domain 2 | ThermoShift | BRD |
BRD4-2-His | BRD4 | Bromodomain-containing protein 4 domain 2 | ThermoShift; AlphaScreen | BRD |
BRD4-Tndm-GST | BRD4 | Bromodomain-containing protein 4 domain 1 and 2 | ThermoShift | BRD |
BRD4-Tndm-His | BRD4 | Bromodomain-containing protein 4 domain 1 and 2 | ThermoShift; AlphaScreen | BRD |
BRD7-GST | BRD7 | BP75; CELTIX-1 | ThermoShift | BRD |
BRD9-GST | BRD9 | Bromodomain-containing protein 9 | ThermoShift | BRD |
BRD9-His | BRD9 | Bromodomain-containing protein 9 | ThermoShift; AlphaScreen | BRD |
BRDT-1-His | BRDT | Bromodomain testis-specific protein domain 1 | ThermoShift; AlphaScreen | BRD |
BRDT-2-His | BRDT | Bromodomain testis-specific protein domain 2 | ThermoShift | BRD |
BRDT-Tndm-His | BRDT | Bromodomain testis-specific protein domain 1 and domain 2 | ThermoShift; AlphaScreen | BRD |
BRPF1a | BRPF1 | Bromodomain and PHD finger-containing protein 1, isoform 1 | ThermoShift | BRD |
BRPF1b-GST | BRPF1 | Bromodomain and PHD finger-containing protein 1, isoform 2 | ThermoShift | BRD |
BRPF1b-His | BRPF1 | Bromodomain and PHD finger-containing protein 1, isoform 2 | ThermoShift; AlphaScreen | BRD |
BRPF3-GST | BRPF3 | Bromodomain and PHD finger-containing protein 3 | ThermoShift | BRD |
BRPF3-His | BRPF3 | Bromodomain and PHD finger-containing protein 3 | ThermoShift; AlphaScreen | BRD |
BRWD1-2-GST | BRWD1 | Bromodomain and WD repeat-containing protein 1-Bromodomain 2 | ThermoShift | BRD |
BRWD1-2-His | BRWD1 | Bromodomain and WD repeat-containing protein 1-Bromodomain 2 | ThermoShift; AlphaScreen | BRD |
BRWD3-2-GST | BRWD3 | Bromodomain and WD repeat-containing protein 3-Bromodomain 2 | ThermoShift | BRD |
CBX7-[CHR]-GST | CBX7 | Chromobox 7 chromodomain | ThermoShift | CHR |
CDg1-[CHR]-GST | CDY1 | chromodomain Y-linked 1 chromodomain | ThermoShift | CHR |
CECR2-GST | CECR2 | Cat eye syndrome critical region protein 2 | ThermoShift | BRD |
CECR2-His | CECR2 | Cat eye syndrome critical region protein 2 | AlphaScreen; ThermoShift | BRD |
CHD1-[CHR]-GST | CHD1 | chromodomain helicase DNA binding protein 1 chromodomain | ThermoShift | CHR |
CHD1-[CHR]-His | CHD1 | chromodomain helicase DNA binding protein 1 chromodomain | ThermoShift | CHR |
CHD2-[CHR]-GST | CHD2 | chromodomain helicase DNA binding protein 2 chromodomain | ThermoShift | CHR |
CHD2-[CHR]-His | CHD2 | chromodomain helicase DNA binding protein 2 chromodomain | ThermoShift | CHR |
CHD4-[CHR]-GST | CHD4 | chromodomain helicase DNA binding protein 4 chromodomain | ThermoShift | CHR |
CHD4-[PHD-CHR]-GST | CHD4 | chromodomain helicase DNA binding protein 4 PHD domain and chromodomain | ThermoShift | CHR; PHD |
CHD7-[CHR]-GST | CHD7 | chromodomain helicase DNA binding protein 4 chromodomain | ThermoShift | CHR |
CREBBP-GST | CREBBP | CREB-binding protein | ThermoShift | BRD |
CREBBP-His | CREBBP | CREB-binding protein | ThermoShift; AlphaScreen | BRD |
EED | EED | Polycomb protein EED, embryonic ectoderm development | HTRF | WDR |
EP300-GST | EP300 | Histone acetyltransferase p300, E1A-associated protein p300 | ThermoShift | BRD |
EP300-His | EP300 | Histone acetyltransferase p300, E1A-associated protein p300 | ThermoShift | BRD |
HP1alpha-[CHR]-His | CBX5 | HP1a, Chromobox 5 chromodomain | ThermoShift | CHR |
HP1alpha-GST | CBX5 | HP1a, Chromobox 5 | ThermoShift | CHR |
HP1beta-[CHR]-GST | CBX1 | HP1b, Chromobox 1 chromodomain | ThermoShift | CHR |
HP1beta-[CHR]-His | CBX1 | HP1b, Chromobox 1 chromodomain | AlphaScreen*; ThermoShift | CHR |
HP1beta-GST | CBX1 | HP1b, Chromobox 1 | ThermoShift | CHR |
HP1beta-His | CBX1 | HP1b, Chromobox 1 | AlphaScreen*; ThermoShift | CHR |
HP1beta-Strep | CBX1 | HP1b, Chromobox 1 | ThermoShift | CHR |
HP1gamma-GST | CBX3 | Chromobox 3 | ThermoShift | CHR |
HP1gamma-His | CBX3 | Chromobox 3 | ThermoShift | CHR |
KAT2A | KAT2A | lysine acetyltransferase 2A | ThermoShift | BRD |
KAT2B | KAT2B | lysine acetyltransferase 2B | ThermoShift | BRD |
KAT5-2-[CHR]-His | KAT5 | lysine acetyltransferase 5 transcript variant 2 chromodomain | ThermoShift | CHR-like |
KAT5-3-[CHR]-GST | KAT5 | lysine acetyltransferase 5 transcript variant 3 chromodomain | ThermoShift | CHR-like |
KAT5-3-[CHR]-His | KAT5 | lysine acetyltransferase 5 transcript variant 3 chromodomain | ThermoShift | CHR-like |
L3MBTL1 | L3MBTL1 | lethal(3)malignant brain tumor-like protein 1 | ThermoShift | MBT |
L3MBTL1-His | L3MBTL1 | lethal(3)malignant brain tumor-like protein 1 | AlphaScreen | MBT |
MPP8-[CHR]-GST | MPHOSPH8 | M-phase phosphoprotein 8 chromodomain | ThermoShift | CHR |
PB1-1 | PBRM1 | polybromo 1, PBRM1 bromodomain 1 | ThermoShift | BRD |
PB1-2 | PBRM1 | polybromo 1, PBRM1 bromodomain 2 | ThermoShift | BRD |
PB1-3 | PBRM1 | polybromo 1, PBRM1 bromodomain 3 | ThermoShift | BRD |
PB1-4 | PBRM1 | polybromo 1, PBRM1 bromodomain 4 | ThermoShift | BRD |
PB1-5 | PBRM1 | polybromo 1, PBRM1 bromodomain 5 | ThermoShift | BRD |
PB1-6 | PBRM1 | polybromo 1, PBRM1 bromodomain 6 | ThermoShift | BRD |
PHIP-2 | PHIP | pleckstrin homology domain interacting protein bromodomain 2 | ThermoShift | BRD |
PHIP-Tndm | PHIP | pleckstrin homology domain interacting protein tandem bromodomains | ThermoShift | BRD |
SMARCA2a bromodomain | SMARCA2 | SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 2 bromodomain | HTRF | BRD |
SMARCA2a-His | SMARCA2 | SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 2 | ThermoShift | BRD |
SMARCA2b-His | SMARCA2 | SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 2 | AlphaScreen; ThermoShift | BRD |
SMARCA4-His | SMARCA4 | SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4 | ThermoShift; AlphaScreen | BRD |
SP100-GST | SP100 | SP100 nuclear antigen | ThermoShift | BRD |
SP100-His | SP100 | SP100 nuclear antigen | ThermoShift | BRD |
SP110c-GST | SP110 | SP110 nuclear body protein | ThermoShift | BRD |
SP140-GST | SP140 | SP140 nuclear body protein | ThermoShift | BRD |
SP140-His | SP140 | SP140 nuclear body protein | ThermoShift; AlphaScreen | BRD |
SP140L-GST | SP140L | SP140 nuclear body protein like | ThermoShift | BRD |
SP140L-His | SP140L | SP140 nuclear body protein like | ThermoShift; AlphaScreen | BRD |
TAF1-1-GST | TAF1 | TATA-box binding protein associated factor 1, TBP-associated factor, bromodomain 1 | ThermoShift | BRD |
TAF1-2-GST | TAF1 | TATA-box binding protein associated factor 1, TBP-associated factor, bromodomain 2 | ThermoShift | BRD |
TAF1-2-His | TAF1 | TATA-box binding protein associated factor 1, TBP-associated factor, bromodomain 2 | AlphaScreen | BRD |
TAF1L-1 | TAF1L | TAF1-like bromodomain 1 | ThermoShift | BRD |
TAF1L-2 | TAF1L | TAF1-like bromodomain 2 | ThermoShift | BRD |
TAF1L-Tndm-GST | TAF1L | TAF1-like tandem bromodomains | ThermoShift | BRD |
TRIM24 | TRIM24 | tripartite motif containing 24 | ThermoShift | BRD |
TRIM28 | TRIM28 | tripartite motif containing 28 | ThermoShift | BRD |
TRIM33a | TRIM33 | tripartite motif containing 33 transcript variant a | ThermoShift | BRD |
TRIM33b-His | TRIM33 | tripartite motif containing 33 transcript variant a | ThermoShift | BRD |
TRIM66 | TRIM66 | tripartite motif containing 66 | ThermoShift | BRD |
UHRF1 Full length | UHRF1 | ubiquitin like with PHD and ring finger domains 1 full length | ThermoShift | PHD; TDR; SRA |
UHRF1-[PHD] | UHRF1 | ubiquitin like with PHD and ring finger domains 1 PHD | ThermoShift | PHD |
UHRF1-[PHD]-His | UHRF1 | ubiquitin like with PHD and ring finger domains 1 PHD | AlphaScreen* | PHD |
UHRF1-[SRA] | UHRF1 | ubiquitin like with PHD and ring finger domains 1 SRA dmain | ThermoShift | SRA |
UHRF1-[TDR-PHD] | UHRF1 | ubiquitin like with PHD and ring finger domains 1 tudor domain and PHD | ThermoShift | PHD; TDR |
UHRF1-[TDR-PHD]-His | UHRF1 | ubiquitin like with PHD and ring finger domains 1 tudor domain and PHD | AlphaScreen* | PHD; TDR |
UHRF1-[TDR]-His | UHRF1 | ubiquitin like with PHD and ring finger domains 1 tudor domain | AlphaScreen*; ThermoShift | TDR |
UHRF1-His Full length | UHRF1 | ubiquitin like with PHD and ring finger domains 1 full length | AlphaScreen* | PHD; TDR; SRA |
L3MBTL3-His | L3MBTL3 | lethal(3)malignant brain tumor-like protein 3 | AlphaScreen | MBT |
YTHDF1 | YTHDF1 | YTH domain-containing family protein 1, C20orf21, DF1, Dermatomyositis associated with cancer putative autoantigen 1, DACA1 | HTRF | RNA Me |
YTHDF2 | YTHDF2 | HGRG8, CLL-associated antigen KW-14, High-glucose-regulated protein 8, Renal carcinoma antigen NY-REN-2 | HTRF | RNA Me |
* no small molecule control available
BRD... Bromodomain; read acetyl-lysines
CHR ... Chromodomain; read methyl-lysines
PHD ... Plant homeodomain; read methyl-lysines, acetylated-lysines, methyl-arginines, unmodified lysines
MBT ... Malignant brain tumor domain; read methyl- lysines
TDR ... Tudor domain; read methyl-lysines and methyl-arginines
WDR ... WD40-repeat domain; versatile binding
SRA ... SET- and RING-associated domain; read methyl-cytosine
Source: SGC, Chromohub (http://apps.thesgc.org/resources/phylogenetic_trees/)
Assay Details
AlphaScreen assays stand for amplified luminescent proximity homogeneous assay.
Example of bromodomain BRD-1: The His-tagged bromodomain and the substrate, which is a biotin-tagged acetylated peptide, are each captured on AlphaScreen beads. Binding interaction between the peptide and the bromodomain brings the beads in close proximity. Laser excitation of the donor beads allows for energy transfer by singlet oxygen atoms between the fluorophores.
Incubation with test molecules binding to the bromodomain results in a diminished interaction with the biotinylated substrate leading to a decline in light emission.
Thermal shift assays measure the thermal stability of a target protein and the increase in protein melting temperature upon the binding of a ligand to the protein. Our thermal shift bromodomain assays are available as service and as a kit for in-house selectivity screening.
Differential Scanning Fluorimetry of BRD2-Tndm (His) in the Presence or Absence of JQ1. Thermal denaturation of BRD2-Tndm (His) was detected by increased binding and fluorescence of the dye SYPRO Orange. Addition of the BET bromodomain inhibitor JQ1 stabilizes the protein folding and shifts the melting temperature from 41 to 44 degrees Celsius.
HTRF (Homogeneous Time-Resolved Fluorescence) is an assay technology that combines fluorescence resonance energy transfer technology (FRET) with time-resolved measurement (TR) for measuring molecular interactions in a homogeneous format.
HTRF makes use of rare earth complexes such as Europium or Terbium cryptates. These chelates are very stable under a wide range of chemical conditions, are not subject to photobleaching, and show high kinetic stability. HTRF allows enhanced assay performance compared to conventional fluorescent probes in terms of sensitivity, assay window, and robustness.
HTRF assay procedure with a bromodomain: Acetylated peptides are used as substrates for the bromodomains to bind on. Antibodies recognizing the GST tagged-bromodomain are coupled with the donor fluorophore. The peptide is tagged with biotin and recognized by streptavidin which is coupled with an acceptor fluorophore. The proximity of acceptor and donor fluorophore allows for FRET upon excitation.
Incubation with test molecules binding to the bromodomain results in a diminished interaction with the biotinylated substrate leading to a decline in light emission.
Setups: Single-dose screening in duplicates or IC50 value determination with 5 or 10 concentrations. Other screening formats are available upon request.
The thermal shift assay format allows a 'yes' or 'no' answer whether the test molecule binds to the reader domain protein. IC50 determination is not possible.
Controls: No inhibitor (DMSO vehicle) control and for every assay, one target-specific control compound is tested in 10-dose IC50 format.
Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.
Report: The raw data, % inhibition and control compound IC50 values will be reported in Excel format for single-dose assays. For IC50 orders, raw data, IC50 values, and curve fitting will be delivered in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.
Screening facility: This assay is performed at our screening facility in Malvern, PA, US.
Compound requirements: In brief, for a standard project, 20 µl of a 10 mM DMSO stock or solid material is needed. Less material is needed for large-scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.
On a monthly basis, Reaction Biology performs compound screening on the full bromodomain panel.
Please see the upcoming screening dates here.
For more information about the selectivity screening of bromodomain proteins visit our BromoMELTTM Assay Kit page
We offer two assay formats for intracellular compound screening on bromodomain targets:
1. Binding of a compound to BET Bromodomain in intact cells based on NanoBRET Intracellular Target Engagement assay technology.
2. Testing the protein-protein interaction of bromodomains with acetylated histones in intact cells based on the NanoBRET Bromodomain/Histone Interaction Assay.
Please see more information on our Cell-based Epigenetic Assays webpage.