Products from BPS Bioscience require a minimum order value above 400€Application: Identify and optimize Molecular Glues/PROTACs targeting CDK12/Cyclin K or CDK13/Cyclin K and any DDB1-containing E3 ligase complex.Design novel molecules targeting CDK12(13)/Cyclin K and DDB1.Directly compare the activity of different Molecular Glues/PROTACs.
Background: CDKs (cyclin-dependent kinases) are serine/threonine kinases involved in a myriad of critical celullar functions. CDK12 (cyclin-dependent kinase 12)/Cyclin K plays a critical pathophysiological role in cancer and other diseases through its involvement in transcription regulation, DNA damage repair, and genomic stability. CDK12/13 regulate transcription by phosphorylating the C-terminal domain of RNA polymerase II on Ser2, regulating the expression of DNA damage response (DDR) proteins. Understanding the mechanisms underlying CDK12/Cyclin K and how its inactivation can cause genomic instability, can lead to the development of novel targeted therapies and personalized treatment strategies for patients. DDB1 (DNA damage-binding protein 1) functions as a core component of the Cullin 4 (CUL4)-based E3 ubiquitin ligase complexes. DDB1 serves as a bridge or adaptor protein which interacts with Cereblon or with DCAFs (DDB1 and CUL4-associated factors), which are ubiquitin ligase substrates. Molecular glues such as (R)-CR8 induce the formation of a complex between CDK12/Cyclin K and the CUL4 adaptor protein DDB1, directly presenting CDK12/cyclin K for ubiquitination and degradation. CR8 is considered a pan-CDK inhibitor, by acting as a monovalent degrader of cyclin K. CR8 is derived from seliciclib, a CDK inhibitor with no solvent-exposed pyridine. The introduction of solvent exposure pyridine in CR8 created a new mechanism of action, highlighting the real impact of small molecular structure modifications to create innovative tools. The development of these molecular glues and PROTACS opens new avenues in cancer therapy.
Contraindications: The final concentration of DMSO in the assay should not exceed 1%.Avoid green and blue dyes that absorb light in the AlphaScreen signal emission range (λ=520-620 nm), such as Trypan Blue.Avoid the use of potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+).The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. Media like MEM, which lacks these components, does not affect AlphaScreen assays.
Description: The Molecular Glue/PROTAC® Optimization Kit for CDK/Cyclin K-DDB1 Containing Complex is an AlphaLISA® assay designed for the testing and profiling of Molecular Glues (MG) and PROTACs targeting CDK12 (cyclin dependent kinase 12) or CDK13 kinases and any complex containing DDB1 (DNA damage binding protein 1). The kit comes with enough recombinant CDK12/Cyclin K complex, Cereblon/DDB1/CUL4A (culin 4A)/Rbx1 (RING-box protein 1) as a DDB1-containing complex, and optimized PROTAC® buffer. This kit also contains the control molecular glue (R)-CR8, and the dual CDK12-CDK13 inhibitor THZ531.
Storage Stability:
This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Uniprot: Cereblon: Q96SW2; DDB1: Q16531; Cul4a: Q13619; Rbx1: P62877
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References:
Slabicki M., et al., 2020 Nature 585(7824): 293-297.
Lv Lu, et al., 2020 eLife; 9:e59994.
Thomas K., et al., 2024 ACS Chem Biol 19(1):173-184.
More Information
| SKU |
BPS82588 |
| Manufacturer |
BPS Bioscience |
| Manufacturer SKU |
82588 |
| Package Unit |
384 reactions |
| Quantity Unit |
PAK |
| Application |
Dot Blot |
| Product information (PDF) |
Download |
| MSDS (PDF) |
|
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