REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Mouse monoclonal anti PFTAIRE-1 (C-3) | Santa Cruz Biotechnology | Cat # sc-376366 |
Mouse monoclonal anti Flag | Sigma Aldrich | Cat # F1804-50UG |
Rabbit polyclonal anti Cyclin Y | Bethyl Laboratories | Cat # A302-376A |
Rabbit polyclonal anti PCTAIRE-1 | Cell Signaling Technologies | Cat # 4852 |
Rabbit polyclonal anti PCTAIRE-2 | Sigma Aldrich | Cat # SAB1306378 |
Rabbit monoclonal anti CDK2 | Cell Signaling Technologies | Cat # 2546 |
Rabbit monoclonal anti CDK9 | Cell Signaling Technologies | Cat # 2316 |
Rabbit monoclonal anti CDK10 | Cell Signaling Technologies | Cat # 36106 |
Rabbit polyclonal anti Cyclin K | Bethyl Laboratories | Cat # A301-939A |
Mouse monoclonal anti CDK5 | Cell Signaling Technologies | Cat # 12134 |
Rabbit monoclonal anti GSK3α | Cell Signaling Technologies | Cat # 4337 |
Rabbit monoclonal anti Phospho-LRP6 S1490 | Cell Signaling Technologies | Cat # 2568 |
Rabbit monoclonal anti LRP6 | Cell Signaling Technologies | Cat # 2560 |
Rat monoclonal anti Pol II pSer2 | EMD Millipore | Cat # 04-1571 |
Rabbit polyclonal anti Pol II (N-20) | Santa Cruz Biotechnology | Cat # sc-899 |
Rabbit polyclonal anti Phospho-NPM T199 | Cell Signaling Technologies | Cat # 3451 |
Rabbit polyclonal anti NPM | Cell Signaling Technologies | Cat # 3452 |
Rabbit monoclonal anti Phospho-RB S608 | Cell Signaling Technologies | Cat # 8147 |
Rabbit monoclonal anti Phospho-RB S780 | Cell Signaling Technologies | Cat # 8180 |
Rabbit polyclonal anti Phospho-RB S795 | Cell Signaling Technologies | Cat # 9301 |
Rabbit monoclonal anti Phospho-RB S807/811 | Cell Signaling Technologies | Cat # 8516 |
Mouse monoclonal anti RB | Cell Signaling Technologies | Cat # 9309 |
Rabbit monoclonal anti active β-catenin | Cell Signaling Technologies | Cat # 8814 |
Rabbit monoclonal anti β-catenin | Cell Signaling Technologies | Cat # 9582 |
Rabbit polyclonal anti Cyclin B1 | Cell Signaling Technologies | Cat # 4138 |
Rabbit monoclonal anti Phospho-Histone H3 S10 | Cell Signaling Technologies | Cat # 53348 |
Mouse monoclonal anti Histone H3 | Cell Signaling Technologies | Cat # 14269 |
Mouse monoclonal anti Tubulin | Cell Signaling Technologies | Cat # 3873 |
Mouse monoclonal anti Actin | Cell Signaling Technologies | Cat # 3700 |
Bacterial and Virus Strains | ||
NEB 10-beta | NEB | Cat # C3019I |
NEB Stable | NEB | Cat # C3040I |
Biological Samples | ||
Chemicals, Peptides, and Recombinant Proteins | ||
CDK14 / Cyclin Y | Life Technologies | Cat # PV6382 |
CDK14 / Cyclin Y | Signal Chem | Cat # P15-10G-05 |
CDK16 / Cyclin Y | Life Technologies | Cat # PV6379 |
CDK18 / Cyclin Y | Signal Chem | Cat # P11-10G-05 |
RB | Signal Chem | Cat # R05-55G-50 |
CDK2 / Cyclin A | Signal Chem | Cat # C29-18G-10 |
CDK3 / Cyclin A | Signal Chem | Cat # C30-10G-10 |
CellTiter Glo | Promega | Cat # G7571 |
Thymidine | Sigma Aldrich | Cat # T1895-5G |
Peptide: RbS608 TAADMYLSPVRSPKK |
Tufts University Dept. of Physiology | Custom synthesis |
Peptide: RbS780 STRPPTLSPIPHIPR |
Tufts University Dept. of Physiology | Custom synthesis |
Peptide: CDC20 KEAAGPAPSPMRAAN |
Tufts University Dept. of Physiology | Custom synthesis |
Peptide: CCAR2 NQPLLKSPAPPLLH |
Tufts University Dept. of Physiology | Custom synthesis |
Peptide: CENPF KIFTTPLTPSQYYSG |
Tufts University Dept. of Physiology | Custom synthesis |
Peptide: ERF EAGGPLTPRRVSS |
Tufts University Dept. of Physiology | Custom synthesis |
Critical Commercial Assays | ||
LanthaScreen Eu (CDK14 / Cyclin Y) | Invitrogen | Assay ID 1839 |
LanthaScreen Eu (CDK16 / Cyclin Y) | Invitrogen | Assay ID 1838 |
Z’LYTE (CDK2 / Cyclin A) | Invitrogen | Assay ID 314 |
CDK14 33P Kinase Assay | Reaction Biology Corp | |
Deposited Data | ||
Experimental Models: Cell Lines | ||
HCT116 | ATCC | CCL-247 |
HEK293 | ATCC | CRL-1573 |
MDA-MB-231 | ATCC | HTB-26 |
HepG2 | ATCC | HB-8065 |
PATU-8988T | DSMZ | Cat # ACC 162 |
Experimental Models: Organisms/Strains | ||
Oligonucleotides | ||
CDK14 C218S QuikChange forward: GTGCACACTGATTTATCTCAGTACATGGAC | IDT | Custom synthesis |
CDK14 C218S QuikChange reverse: GTCCATGTACTGAGATAAATCAGTGTGCAC | IDT | Custom synthesis |
CDK14 QuikChange sequencing: GAAGAAGAAGGGACAC | IDT | Custom synthesis |
sgCDK14 gRNA forward: caccgTCATGACATCATCCATACCA | IDT | Custom synthesis |
sgCDK14 gRNA reverse: aaacTGGTATGGATGATGTCATGAc | IDT | Custom synthesis |
Recombinant DNA | ||
pCMV6-CDK14-myc/flag | Origene | Cat # RC224426 |
pCMV6-CDK14 C218S-myc/flag | This paper | n/a |
pX458 | Addgene | Cat # 48138 |
pX458-sgCDK14 | This paper | n/a |
pCDH-CMV-MCS- | System Biosciences | Cat # CD510B-1 |
NanoBRET CDK2 | Promega | n/a |
NanoBRET CDK14 | Promega | n/a |
NanoBRET CDK15 | Promega | n/a |
NanoBRET CDK16 | Promega | n/a |
NanoBRET CDK17 | Promega | n/a |
NanoBRET CDK18 | Promega | n/a |
NanoBRET Cyclin Y (carrier DNA) | Promega | n/a |
Software and Algorithms | ||
Prism | GraphPad | |
FlowJo | FlowJo, LLC | |
ImageJ | NIH | https://imagej.nih.gov/ij/download.html |
ZEN lite | Carl Zeiss | https://www.zeiss.com/microscopy/us/products/microscope-software/zen-lite.html |
TIDE | Brinkman et al., 2014 | https://tide.deskgen.com/ |
GO | Ashburner et al., 2000 | http://geneontology.org/page/go-enrichment-analysis |
R - Bioconductor | Huber et al., 2015 | https://www.bioconductor.org/packages/release/bioc/html/limma.html |
Other | ||
Streptavidin-agarose beads | Thermo Fisher | Cat # 20349 |
ProLong Gold DAPI Antifade | Cell Signaling Technologies | Cat # 8961 |
Phalloidin CruzFluor-488 | Santa Cruz | Cat # sc-363791 |