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PRODUCTS and SERVICES Drug Discovery Tools from Carna Biosciences ~Your Kinase Company~ Kinases ~Available from Carna Biosciences~ PRODUCTS AND SERVICES Drug Discovery Tools from Carna Biosciences ~your kinase company~ ~Available from Carna Biosciences~ Kinases Mutants ABL [E255K] KIT [D816E] ABL [T315I] KIT [D816V] ALK [C1156Y] KIT [D816Y] ALK [F1174L] KIT [T670I] Kinome Wide Coverage! ALK [G1202R] KIT [T670I/D816V] ALK [G1269A] KIT [V560G] ALK [L1196M] KIT [V560G/D816V] ALK [L1196M/G1202R] KIT [V654A/D816V] PDGFRβ PDGFRα ALK [R1275Q] LRRK2 [G2019S] FMS HER2 MLK1 TYRO3 EGFR ALK [T1151_L1152insT] MAP2K1 [F129L] KIT MLK4 MLK2 MER AXL HER4 LZK DLK ALK2 [Q207D] MAP2K1 [P124L] FLT3 RON ALK2 [R206H] MET [D1228H] KDR RYK HER3/ErbB3 FLT4 MET TYK2 MLK3 TAK1-TAB1 TKL BRAF [T599 V600insT] MET [D1228N] TK FLT1 TIE1 JAK3 FGFR4 JAK1 ZAK HH498 LIMK1 IRAK1 BRAF [V600E] MET [M1250T] JAK2 TESK2 FGFR1 TIE2 TNK1 LIMK2 IRAK3 FGFR3 ACK BTK [C481S] MET [Y1230C] IGF1R CYGD PBK TESK1 IRR FGFR2 RET LMR1 CYGF IRAK2 ACVR2A BTK [T316A] MET [Y1230H] LMR2 ANPa MOS INSR FER FES ANPb ACVR2B BTK [T474I] MET [Y1235D] LTK LMR3 ARAF HSER IRAK4 TGFβR2 BTK [T474S] NPM1-ALK MISR2 ALK ROS BRAF RAF1 ALK2 EGFR [C797S] PDGFRa [D842V] BTK ILK BMPR2 ALK1 KSR2 BMPR1A EGFR [C797S/L858R] PDGFRa [T674I] BMX ITK EphA10 SuRTK106 CCK4 MLKL BMPR1B EGFR [d746-750] PDGFRa [V561D] TXK CSK KSR1 EphB6 PYK2 TGFβR1 EGFR [d746-750/C797S] RET [G691S] TEC CTK FAK ZAP70 ALK7 ALK4 ANKRD3 EGFR [d746-750/T790M] RET [M918T] SRM SYK SgK288 EphA1 ROR2 RIPK3 RIPK2 EGFR [d746-750/T790M/C797S] RET [S891A] BRK ROR1 ARG ABL MUSK RIPK1 EGFR[D770_N771 ins NPG] RET [Y791F] FRK EphA2 EGFR [L858R] RET[V804M] BLK LRRK2 DDR1 DDR2 TRKA SgK496 LRRK1 EGFR [L861Q] YES(YESs) [T348I] LCK EphA8 EGFR [T790M] HCK TRKC SRLK6 MST3 EGFR [T790M/C797S/L858R] LYNa, LYNb FGR TRKB SRLK5 MST1 YSK1 EGFR [T790M/L858R] SRC FYN PAK1 EphA6 EphB4 SgK307 MST2 MST4 EML4-ALK [isoform a] PAK3 PAK4 YES [isoform b] EphB1 SgK071 TAO1 FGFR1 [V561M] EphB3 Slob PAK5 EphA7 HIPK1 TAOK2 FGFR2 [N549H] EphB2 PAK2 PAK6 HIPK2 HIPK3 HGK FGFR2 [V564I] EphA4 IRE1 TBCK TAOK3 EphA3 IRE2 SCYL2 MINK SCYL1 FGFR3 [G697C] EphA5 MSSK1 STLK3 FGFR3 [K650E] DYRK1A HIPK4 SRPK1 OSR1 TNIK DYRK1B SRPK2 SLK ZC4/NRK FGFR3 [K650M] LOK MYO3A KHS1 DYRK4 RNAseL MYO3B KHS2 FGFR3 [V555L] KIS CAMK2δ CAMK PRP4 PRPK MAP4K2 FGFR3 [V555M] β DYRK2 HPK1 MAP3K3 CAMK2α CAMK2 MAP3K2 DYRK3 CLK3 YSK4 FGFR4 [N535K] γ FGFR4 [V550E] PHKG1 CAMK2 CLK2 MAP3K1 MAP3K4 PHKG2 MAPKAPK2 MAP3K14 MAP3K5 FGFR4 [V550L] DCAMKL2 CASK TSSK2 TSSK1 CLK4 MAP3K6 MAPKAPK3 COT MAP3K15 JAK2 (JH1 JH2)[V617F] DCAMKL1 DCAMKL3 CLK1 MAP2K2 MAP2K5 PSKH2 MNK1 PIM3 MAP2K7 MAP2K1 PSKH1 MAPKAPK5 TSSK3 CAMK4 VACAMKL CHK2 MNK2 PIM1 CAMK1β WNK1 WNK3 MAP2K4 CAMK1g PIM2 SSTK CaMKK2 CAMK1α NRBP1 MAP2K6 MGC42105 CaMKK1 NRBP2 δ TSSK4 MAP2K3 STE CAMK1 NuaK2 SNRK PASK LKB1 BIKE WNK2 BRSK2 NuaK1 HUNK AAK1 WNK4 BRSK1 CHK1 CDC7 MELK GAK SCYL3 AMPKα2/β1/γ1, AMPKα2/β2/γ1 Qsk PIK3R4 SIK CLIK1 MPSK1 AMPKα1/β1/γ1, AMPKα1/β2/γ1 QIK CLIK1L BUB1 Trad SPEG.d2 Obscn TLK1 SgK396 BUBR1 MARK4 δ Trio TTK TLK2 CK1 MARK2 SPEG.d1 STK33 CK1ε DRAK2 Obscn.d1 Haspin α MARK1 CK1 Atypical DRAK1 TTN CK1a2 CK1 DAPK2 VRK3 MARK3 SgK493 CK1γ1 DAPK1 SgK196 TTBK1 γ skMLCK CK1γ2 CK1 3 CHAK1 DAPK3 smMLCK TTBK2 AKT1 AKT2 IKKε VRK1 caMLCK VRK2 PKN2 PKCα EEF2K TBK1 PKD2 YANK1 SGK3 η SgK086 PKN1 PKC PKCβ1 PDHK1 IKKβ YANK2AKT3 SGK PKD3 PKCε PKCβ2 PDHK2 IKKα ULK3 SBK YANK3 PKD1 NEK7 PKN3 PKCγ ULK4 Fused SgK494 SGK2 PDHK3 ULK1 NEK6 MASTL PKCδ ULK2 SgK069 PDHK4 NEK9 SgK110 PKCι PKCθ NEK8 ζ HRI PLK4 AurA PKC NEK2 SgK223 PINK1 PDK1 RSK1 RSK2 NEK10 AurB NEK11 MYT1 SgK269 RSK4 Inactive & GCN2 AurC RSK3 NEK4 WEE1b PLK1 MSK2 WEE1 Inactive-mutants NEK3 PLK2 SgK495 RSKL2 BARK1 MSK1 PKR PLK3 RSKL1 CRIK NEK1 PEK BARK2 p70S6Kb p70S6K CK2α1/β MAST3 Erk2 NEK5 GPRK7 PKACγ PKACβ Erk4 CK2a2/b Trb3 RHOK GPRK6 Erk5 MAST1 ROCK1 PKACα CDK8 GSK3α GPRK5 IKKα(inactive mutant only) Trb1 Trb2 ROCK2 PRKX JNK2 JNK3 Erk7 β GPRK4 MAST4 JNK1 GSK3 MAST2 DMPK1 PRKY JNK1 NDR2 PGK JNK2 (inactive only) Other Erk3 CDKL4 p38β CDKL5 CDKL1 LATS1 NDR1 CGK2 MAP2K1 α CDK11 MOK CDKL3 LATS2 DMPK2 β p38 CDK6 ICK CDKL2 MRCK MAP2K6 γ MAK MRCKα p38 δ NLK CDK4 MAP2K7 p38 AGC Erk5 CDK2 p38a CCRK CDK7 CDK5 CDK3 Erk2 CDC2 Erk1 PITSLRE PCTAIRE3 CDK10 CDK9 PCTAIRE2 Lipid Kinases CMGC PFTAIRE2 CDK9/CycK PFTAIRE1 CRK7 CHED PCTAIRE1(CDK16)/CycY MTOR(FRAP) / MLST8 PIP5K1A PI4KB PIP5K1B PIK3C3 PIP5K1C PIK3CA / PIK3R1 PIP5KL1 PIK3CA[E542K] / PIK3R1 SPHK1 PIK3CA[E545K] / PIK3R1 SPHK2 PIK3CA[H1047R] / PIK3R1 PIK3CA[P539R] / PIK3R1 PIK3CA[R88Q] / PIK3R1 PIK3CB / PIK3R1 PIK3CD / PIK3R1 PIK3CG PIKFYVE(PIP5K3) PIP4K2A PIP4K2B PIP4K2C www.carnabio.com 1 High Quality Active Kinases Carna offers an expanded portfolio of high purity, active human kinases and related products. All of our >450 kinases and products are developed and produced entirely in-house from gene cloning, expression and purification, and undergo rigorous quality control. All products are available in various sizes starting from 5μg up to bulk production, are delivered to you with a lot specific data sheet. *Target lists: Please refer to the attachment. Industry Leading Production and Quality Control Methods Proprietary Production Methods ensure batch-to-batch consistency (Fig.1) Relevant kinase specific substrate(s) measure activity of each batch Target kinase activity maximized to minimize literature documented host cell activity Kinase constructs are carefully selected from published literature DNA sequence confirmed prior to protein expression Amino acid sequence confirmed by Peptide Mass Fingerprinting(PMF) Unparalleled Kinase Activity Production process yields highly active kinases. Some kinases are activated further by : ◆ Expression with upstream kinase(s) ◆ Tag removal ◆ ATP treatment Batch-to-Batch Consistency 1 0.9 0.8 0.7 ) 0.6 S + 0.5 P ( 13CBS-0004 / 0.4 P 0.3 13CBS-0005 0.2 11CBS-1108 0.1 0 0 2 4 6 8 10 12 EGFR (ng/µl) [Fig.1] Lipid Kinases and Related Products Carna’s lipid kinase panel includes numerous members of the PIK family. The enzymes enable complete and easy investigation of lipid kinase drug targets. Protein Substrates Our protein substrates are available for use in phosphoryla- tion activity assays to assess kinase activity. Custom protein production We offer custom production services using our extensive expertise, for any kinase of interest. For more information, please contact us at [email protected] www.carnabio.com 2 Biotinylated Kinases Biotinylated Kinases are best suited for the study of compound binding affinity and other kinase-molecule interactions using devic- es measuring Surface Plasmon Resonance (SPR), BioLayer Interferometry (BLI) and other similar biomolecular interactions. They can also be utilized in homogenous proximity-based binding assays such as TR-FRET, AlphaScreenTM and HTRF® to interrogate inhibitor binding affinity, determine on-off rates, and measure binding kinetics. The immobilization of target proteins onto sensor surfaces without impairing their structure and activity can be challenging in small molecule drug discovery. Carna' s in-house, single-site specifically biotinylated kinases are easily immobilized, leading to rapid acquisition of accurate and real-time data for evaluation of your drug candidates! *Target lists: Please refer to the attachment. Advantages of Carna’s Biological Biotinylation Process Kinases are labeled with a single biotin at the N-terminus Easy-to-use ; no additional labeling required Native, catalytically active kinase domains are preserved High quality human kinases produced via Baculovirus expression system Stable activity determined post expression Select kinases available pre-activated (via ATP treatment) and non-activated (without ATP treatment) Biological Biotinylation Biotin acceptor peptide Baculovirus N-terminus Tag Kinase Biotin Biotin Insect Cell SPR data samples* using Carna’s biotinylated kinases are shown on our website （*Measured using Biacore T200 in collaboration with NTRC） Binding kinetics of bosutinib to ABL Click! Response Time Binding kinetics of ibrutinib to BTK[T474I] BTK[T474I] Ibrutinib Response Time s www.carnabio.com 3 QuickScout Screening Assist™ Kits QuickScout Screening Assist™ Kits are designed to accelerate your in house compound screening, particularly secondary and counter- screening applications. Our Kits provide essential reagents and detailed assay protocols in one package, and are available for more than 300 human kinase targets. After your initial kit purchase, components contained in the kit can be purchased separately and in bulk. *Target lists: Please refer to the attachment. Advantages of Carna's Assay Kits Prepared utilizing the extensive expertise of our kinase profiling team Ready-To-Run products & protocols save time and money Scalable for HTS applications Designed for primary, in-house screening procedures applicable to Lead Generation through Lead Optimization! Assay Platform Minimum Kit Size Kit Components Mobility Shift Assay ● Protein Kinase 400dp ● Substrate Mixture（ATP, Cation included） QSS AssistTM MSA ● Assay Buffer Equivalent to ● Termination Buffer This MSA kit works best using LabChip® 1 x 384-well plate technology from PerkinElmer, Inc. ● Assay Protocol（Separation conditions included） 400dp ● Protein Kinase FP(IMAP™) ● Substrate Mixture（ATP, Cation included） TM Equivalent to ● Assay Buffer QSS Assist FP 1 x 384-well plate ● Assay Protocol 400dp ● Protein Kinase TR-FRET ● Substrate Mixture（ATP, Cation included） Equivalent to QSS AssistTM TR-FRET ● Assay Buffer 1 x 384-well plate ● Assay Protocol ● Protein Kinase 100dp ● Substrate Mixture（ATP, Cation included） ELISA ● Assay Buffer TM Equivalent to QSS Assist ELISA 1 x 96-well plate ● Antibody for ELISA (except for TTK & WEE1) ● Assay Protocol ● Protein Kinase ● Substrate Solution TM ADP-Glo 400dp ● Kinase dilution Buffer（Cation included） ● Assay Buffer QSS AssistTM ADP-GloTM Equivalent to 1 x 384-well plate ● MgCl2 solution for detection reagent ● Assay Protocol Sample protocols for all kit platforms are available online. Each kinase kit is made-to-order, with turnaround time of 2-3 weeks.
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