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. 2021 Sep 15;2(4):100822. doi: 10.1016/j.xpro.2021.100822

Table 1.

Summary of individual NanoBRET(TM) Kinase assay components and expected assay outcomes using control inhibitors CC1 or crizotinib

Kinase Product # (promega) Diluent DNA Orientation of Nluc [NanoBRET(TM) tracer K10], nM BRET S:B (BRETTracer / BRETno tracer) CC1 % occupancy, 300 nM St dev Crizotinib % occupancy, 1 uM St dev
LRRK2 NV340A Transfection Carrier C 25 3.7 98 2.0 −9 15.2
MAPK6 NV169A Transfection Carrier N 25 7.4 99 0.8 2 2.4
IRAK3 NV144A Transfection Carrier N 25 4.5 98 0.8 42 4.4
TEK NV215A Transfection Carrier C 25 3.7 90 1.0 37 4.7
TNK1 NV218A Transfection Carrier N 25 10.0 94 0.8 −6 4.1
GAK NV142A Transfection Carrier N 25 14.9 97 0.5 −14 1.5
MAPK4 NV168A Transfection Carrier N 25 26.8 97 1.2 −12 3.4
AAK1 NV100A Transfection Carrier N 25 12.2 99 0.3 −14 0.9
AURKA NV104A Transfection Carrier C 25 6.8 93 1.0 30 7.7
AURKC NV106A Transfection Carrier C 25 10.6 92 1.8 48 4.2
AURKB NV105A Transfection Carrier C 25 10.7 94 1.6 46 5.9
NUAK1 NV183A Transfection Carrier N 25 3.4 98 2.4 36 6.6
LATS2 NV151A Transfection Carrier C 25 5.0 79 7.1 −10 6.2
RPS6KA3 NV200A Transfection Carrier N 25 3.6 97 0.9 −7 7.7
SNF1LK2 NV206A Transfection Carrier N 25 16.9 99 0.5 0 3.4
MYLK2 NV177A Transfection Carrier C 25 6.2 99 0.4 −16 8.3
AXL NV107A Transfection Carrier C 250 2.9 63 3.8 68 1.5
FGFR3 NV136A Transfection Carrier C 250 3.4 84 4.1 14 6.8
FLT3 NV139A Transfection Carrier C 250 3.4 79 3.7 −4 8.6
IGF1R NV326A Transfection Carrier C 250 2.1 78 3.8 18 8.2
INSR NV327A Transfection Carrier C 250 2.9 83 2.5 20 2.1
LIMK2 NV153A Transfection Carrier C 250 4.2 76 3.6 32 9.8
TEC NV214A Transfection Carrier N 250 4.6 78 1.4 −7 6.7
TIE1 NV217A Transfection Carrier C 250 9.9 67 7.2 35 9.9
CLK1 NV113A Transfection Carrier N 250 7.3 98 0.7 0 3.4
SBK3 NV421A Transfection Carrier N 250 4.1 90 4.4 41 5.6
NEK9 NV180A Transfection Carrier N 250 7.9 65 5.2 1 5.4
NEK3 NV179A Transfection Carrier N 250 16.1 44 9.6 −2 14.5
NIM1K NV381A Transfection Carrier C 250 4.6 82 4.9 −3 1.7
STK36 NV433A Transfection Carrier N 250 7.1 89 3.8 −6 6.5
ULK2 NV222A Transfection Carrier N 250 10.5 78 8.0 −1 1.9
ULK3 NV449A Transfection Carrier N 250 23.1 72 5.2 2 6.5
BRSK1 NV249A Transfection Carrier N 250 8.3 78 7.1 −1 4.0
MAP3K10 NV156A Transfection Carrier N 250 4.5 54 3.3 −1 4.4
MAP3K9 NV160A Transfection Carrier N 250 2.9 60 4.3 −1 6.2
MYLK3 NV374A Transfection Carrier C 250 8.5 72 9.1 −8 6.4
PHKG1 NV186A Transfection Carrier N 250 5.9 86 3.1 −3 6.3
STK33 NV211A Transfection Carrier N 250 3.1 90 3.4 −3 3.9
STK4 NV435A Transfection Carrier N 250 4.8 67 2.4 11 7.6
TLK1 NV443A Transfection Carrier C 250 2.8 5 1.9 −5 3.8
FGFR1 NV134A Transfection Carrier C 100 3.3 95 1.8 0 3.8
FGFR2 NV135A Transfection Carrier C 100 3.5 94 0.7 10 1.8
MUSK NV176A Transfection Carrier C 100 3.3 89 4.3 80 2.8
NTRK1 NV181A Transfection Carrier C 100 4.1 68 4.5 71 2.5
RET NV195A Transfection Carrier C 100 6.0 90 1.5 4 4.5
NTRK2 NV182A Transfection Carrier C 100 4.6 80 3.4 85 2.3
TNK2 NV445A Transfection Carrier C 100 6.8 73 4.0 13 6.7
LTK NV154A Transfection Carrier C 100 4.2 71 4.3 87 0.9
BRAF(V600E) NV248A Transfection Carrier C 100 3.9 39 5.8 −4 3.9
IRAK4 NV145A Transfection Carrier C 100 6.5 98 1.4 6 7.0
ITK NV146A Transfection Carrier N 100 3.4 92 1.8 −1 6.2
JAK2 (V617F) NV330A Transfection Carrier C 100 3.3 90 2.8 65 8.3
MAP3K11 NV157A Transfection Carrier N 100 4.6 71 4.9 0 6.0
PTK2 NV192A Transfection Carrier N 100 3.3 88 1.2 40 4.0
PTK6 NV194A Transfection Carrier C 100 3.7 100 0.6 −2 1.2
PTK2B NV193A Transfection Carrier C 100 2.2 91 2.2 47 5.1
BMP2K NV109A Transfection Carrier N 100 8.9 94 1.6 11 6.0
NEK5 NV379A Transfection Carrier N 100 16.2 85 1.2 −6 2.1
STK16 NV209A Transfection Carrier N 100 13.4 85 1.9 −13 6.2
TBK1 NV213A Transfection Carrier N 100 7.5 78 4.2 −7 10.8
ULK1 NV221A Transfection Carrier N 100 14.2 94 1.9 −1 5.5
MAP4K2 NV162A Transfection Carrier N 100 4.5 76 2.0 77 10.7
WEE1 NV223A Transfection Carrier C 100 5.3 91 1.0 −9 2.5
MYLK4 NV375A Transfection Carrier C 100 6.1 99 0.7 −8 8.4
MARK4 NV173A Transfection Carrier N 100 4.4 90 2.0 −15 4.5
PRKAA1 NV410A Transfection Carrier N 100 11.3 92 1.5 −1 4.9
PRKAA2 NV189A Transfection Carrier N 100 12.6 97 0.3 −6 2.2
RPS6KA1 NV198A Transfection Carrier N 100 7.9 89 1.9 −6 2.5
RPS6KA2 NV199A Transfection Carrier N 100 4.7 79 4.2 −7 7.5
RPS6KA4 NV201A Transfection Carrier N 100 6.6 85 0.7 −11 6.0
RPS6KA6 NV202A Transfection Carrier N 100 3.0 89 2.3 −10 7.3
SIK1 NV203A Transfection Carrier N 100 10.2 93 1.6 −4 4.7
CLK4 NV115A Transfection Carrier C 100 2.5 101 1.4 −19 4.6
MAPK8 NV170A Transfection Carrier N 100 5.8 70 7.8 −16 4.2
MAPK9 NV171A Transfection Carrier N 100 9.4 70 6.4 −11 2.1
IKBKE NV143A Transfection Carrier N 100 7.7 74 5.2 21 2.8
LATS1 NV150A Transfection Carrier C 100 5.1 68 5.7 −10 3.9
PRKX NV191A Transfection Carrier C 100 3.6 65 4.9 −14 7.9
CSNK2A2 NV119A Transfection Carrier C 100 5.5 87 3.0 −14 5.8
HIPK4 NV324A Transfection Carrier N 100 7.8 97 0.4 −9 6.1
STK10 NV426A Transfection Carrier C 1000 2.8 80 3.4 35 3.0
FGFR4 NV137A Transfection Carrier C 1000 3.5 68 4.0 3 1.6
MAP4K1 NV161A Transfection Carrier N 1000 3.9 70 4.5 14 4.1
MERTK NV356A Transfection Carrier C 1000 2.2 48 3.9 58 3.3
MET NV175A Transfection Carrier C 1000 5.1 42 8.5 95 1.1
RON NV418A Transfection Carrier C 1000 3.9 32 9.9 86 4.8
TYRO3 NV448A Transfection Carrier C 1000 2.5 47 0.5 49 2.2
LCK NV152A Transfection Carrier C 1000 3.8 78 4.2 36 5.7
LIMK1 NV339A Transfection Carrier C 1000 5.2 69 5.6 29 5.5
EPHA1 NV122A Transfection Carrier C 1000 3.3 74 3.2 75 2.2
EPHA4 NV124A Transfection Carrier C 1000 2.8 33 5.2 53 4.4
EPHA6 NV126A Transfection Carrier C 1000 3.9 38 1.7 64 5.1
EPHA7 NV127A Transfection Carrier C 1000 4.4 51 4.6 38 2.4
EPHB1 NV307A Transfection Carrier C 1000 3.4 65 2.4 68 2.0
EPHB4 NV131A Transfection Carrier C 1000 3.1 53 4.9 62 3.6
FYN NV141A Transfection Carrier C 1000 3.9 65 7.5 −2 12.3
ABL2 NV233A Transfection Carrier N 1000 2.9 72 5.5 43 3.9
BMX NV110A Transfection Carrier C 1000 4.5 60 6.3 7 4.3
BTK N244A Transfection Carrier C 1000 6.8 75 4.4 1 1.3
FER NV133A Transfection Carrier C 1000 2.8 66 4.9 16 7.9
FES NV309A Transfection Carrier C 1000 3.0 43 7.6 1 2.5
JAK3 NV147A Transfection Carrier C 1000 4.7 45 4.8 2 4.4
SRMS NV425A Transfection Carrier N 1000 2.7 18 4.3 0 4.9
TXK NV220A Transfection Carrier C 1000 2.6 60 13.1 18 2.8
CLK2 NV114A Transfection Carrier C 1000 4.1 89 2.2 1 4.7
DYRK1A NV303A Transfection Carrier N 1000 2.6 78 4.4 0 5.3
DYRK1B NV121A Transfection Carrier N 1000 3.3 87 3.6 0 2.1
ERN1 NV132A Transfection Carrier C 1000 3.6 83 1.3 −3 8.1
ERN2 NV308A Transfection Carrier C 1000 5.3 61 7.4 −9 13.9
HIPK2 NV322A Transfection Carrier N 1000 3.3 82 2.8 5 4.4
HIPK3 NV323A Transfection Carrier N 1000 1.9 70 14.3 −4 14.6
ICK NV325A Transfection Carrier N 1000 2.6 47 11.0 3 15.4
CDK1 NV270A CycB1 (NV260A) C 1000 3.0 71 7.1 7 2.9
CDK2 NV278A CycE1 (NV264A) C 1000 12.5 80 4.0 3 1.6
CDK3 NV280A CycE1 (NV264A) C 1000 6.9 85 3.5 6 1.3
CDK4 NV281A CycD3 (NV263A) N 1000 8.2 93 2.2 4 4.2
CDK5 NV112A CDK5R1 (NV282A C 1000 13.3 85 3.6 1 0.8
CDK6 NV284A CycD1 (NV262A) N 1000 4.1 89 2.2 0 3.0
CDK7 NV285A Transfection Carrier N 1000 3.3 83 4.1 32 1.3
CDK9 NV287A CycK (NV266A) N 1000 3.8 68 4.1 4 5.5
CDK10 NV271A CycL2 (NV267A) C 1000 2.9 50 3.7 8 0.5
CDK14 NV272A CycY (NV269A) C 1000 4.3 82 1.3 0 2.4
CDK15 NV273A CycY (NV269A) N 1000 3.5 78 2.1 3 2.8
CDK16 NV274A CycY (NV269A) C 1000 7.2 96 0.6 2 0.4
CDK17 NV275A CycY (NV269A) C 1000 9.6 95 0.8 3 3.6
CDK18 NV276A CycY (NV269A) C 1000 11.4 86 1.4 2 2.1
CDKL2 NV289A Transfection Carrier N 1000 6.7 95 1.3 21 0.7
CDK20 NV279A CycH (NV265A) N 1000 4.3 54 4.9 −11 5.0
CDKL1 NV288A Transfection Carrier N 1000 5.1 64 3.2 2 0.9
CSNK2A1 NV298A Transfection Carrier C 1000 2.8 64 2.6 5 6.7
CDKL3 NV290A Transfection Carrier N 1000 2.2 83 4.5 8 2.0
CDKL5 NV291A Transfection Carrier N 1000 4.6 84 1.3 4 1.8
JNK3 NV148A Transfection Carrier C 1000 3.1 36 3.9 −4 0.8
MAPK11 NV165A Transfection Carrier N 1000 5.4 35 2.3 0 2.6
MAPK14 NV166A Transfection Carrier C 1000 3.7 22 1.5 0 1.7
NLK NV382A Transfection Carrier C 1000 3.2 30 3.4 −1 3.5
NEK11 NV377A Transfection Carrier N 1000 3.9 52 4.2 7 3.6
NEK1 NV376A Transfection Carrier N 1000 5.9 38 1.7 3 4.1
NEK2 NV178A Transfection Carrier N 1000 3.4 46 2.7 6 6.1
NEK4 NV378A Transfection Carrier N 1000 3.8 25 0.8 −12 5.6
PAK4 NV184A Transfection Carrier C 1000 11.1 25 5.5 1 2.4
MAP4K3 NV163A Transfection Carrier N 1000 7.6 50 1.4 8 1.5
STK11 NV208A Transfection Carrier N 1000 3.5 81 2.3 −1 0.9
SLK NV205A Transfection Carrier N 1000 2.5 87 3.8 36 2.9
DAPK2 NV299A Transfection Carrier N 1000 2.5 53 3.4 3 1.6
MAP3K2 NV347A Transfection Carrier C 1000 3.2 47 2.9 6 1.7
PLK2 NV408A Transfection Carrier N 1000 3.0 71 3.2 4 8.5
PLK3 NV409A Transfection Carrier N 1000 3.2 52 0.8 4 1.9
PLK4 NV188A Transfection Carrier N 1000 8.6 52 3.9 4 1.2
STK35 NV432A Transfection Carrier C 1000 3.4 60 5.9 16 3.3
STK17B NV427A Transfection Carrier C 1000 1.9 56 2.5 0 3.8
TLK2 NV444A Transfection Carrier C 1000 3.7 1 3.9 0 3.2
BRSK2 NV111A Transfection Carrier N 1000 15.9 79 2.5 3 2.4
MARK2 NV172A Transfection Carrier N 1000 4.2 76 1.7 4 2.1
MELK NV174A Transfection Carrier N 1000 5.8 43 6.8 −2 2.4
CSNK1A1L NV295A Transfection Carrier N 1000 3.6 19 4.3 −6 2.9
CSNK1D NV296A Transfection Carrier N 1000 3.5 30 2.3 −4 1.4
CSNK1G2 NV118A Transfection Carrier N 1000 3.4 47 1.2 1 2.8
SIK3 NV204A Transfection Carrier N 1000 12.8 62 0.8 0 3.3
SNRK NV424A Transfection Carrier N 1000 4.0 68 3.7 −4 7.7
CAMK1 NV253A Transfection Carrier N 1000 2.4 54 2.5 7 2.6
CAMK2A NV256A Transfection Carrier N 1000 5.8 64 0.7 3 2.3
CAMK2D NV257A Transfection Carrier N 1000 5.7 68 1.3 3 2.2
CHEK2 NV293A Transfection Carrier C 1000 2.4 40 4.4 0 5.0
DCLK3 NV300A Transfection Carrier C 1000 2.4 61 2.7 0 3.4
MKNK2 NV371A Transfection Carrier N 1000 2.6 64 3.0 1 4.6
PHKG2 NV388A Transfection Carrier N 1000 2.7 50 1.4 3 2.5
MAP3K3 NV349A Transfection Carrier C 1000 3.1 36 3.3 4 1.9
RIOK2 NV196A Transfection Carrier N 1000 7.1 31 3.4 9 6.8
MAP4K5 NV350A Transfection Carrier C 1000 3.2 26 1.2 36 4.0
MAST3 NV353A Transfection Carrier N 1000 2.5 44 8.8 −1 6.6
MAST4 NV354A Transfection Carrier N 1000 2.2 46 6.0 6 6.4
STK32B NV210A Transfection Carrier N 1000 3.8 18 4.3 −1 1.9
STK3 NV430A Transfection Carrier N 1000 3.9 43 3.5 1 8.8
STK38 NV212A Transfection Carrier C 1000 6.8 24 0.5 1 2.2
STK38L NV434A Transfection Carrier C 1000 3.9 20 2.1 5 2.8
PAK6 NV386A Transfection Carrier C 1000 3.3 24 13.4 −7 23.7
AKT2 NV103A Transfection Carrier C 1000 3.5 22 1.7 −1 4.7
PKMYT1 NV187A Transfection Carrier C 1000 2.6 30 1.1 6 1.7
PRKACA NV190A Transfection Carrier C 1000 6.9 56 3.1 −1 1.4
PRKACB NV411A Transfection Carrier C 1000 3.6 26 3.5 5 3.5
PRKCE NV412A Transfection Carrier C 1000 3.3 30 8.1 −6 6.1
SGK1 NV422A Transfection Carrier C 1000 3.0 43 4.5 −1 1.1
WEE2 NV450A Transfection Carrier N 1000 2.8 69 2.7 −4 2.6
RIPK1 NV417A Transfection Carrier N 1000 2.8 25 1.8 23 3.9
RIPK2 NV197A Transfection Carrier N 1000 9.1 64 1.5 43 0.5
TNNI3K NV446A Transfection Carrier C 1000 4.0 65 0.9 85 1.4
MLTK NV372A Transfection Carrier N 1000 4.4 62 5.4 −3 8.7
MAP3K12 NV158A Transfection Carrier N 1000 3.7 39 1.0 12 6.8
MAP3K19 NV346A Transfection Carrier C 1000 3.0 8 4.4 6 5.7
MAP3K21 NV348A Transfection Carrier N 1000 3.2 33 0.6 3 5.3
MAP3K4 NV159A Transfection Carrier C 1000 4.8 90 1.7 −1 2.6