PROTAC® Optimization Kit for PARP1-Cereblon Binding

Catalog #

78441

$995 *

Size: 384 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The PROTAC® Optimization Kit for PARP1-Cereblon Binding is designed for the testing and profiling of PROTACs directed against PARP1 (Poly (ADP-ribose)-polymerase 1) and Cereblon. The PROTAC® Optimization Kit for PARP1-Cereblon Binding comes in a convenient AlphaLISA® format, with enough PARP1 Degrader iRucaparib-AP6 PROTAC®, assay buffer, recombinant purified PARP1 and CRBN (Cereblon)/ DDB1(damage specific DNA binding protein 1)/Cul4A (cullin 4A)/ Rbx1 (ring-box 1) complex for 384 reactions. The PARP1 inhibitor Rucaparib is included as a control inhibitor of PROTAC binding to PARP1 (internal control).

Figure 1. Schematic representation of PARP1-Cereblon complex formation via iRucaparib-AP6 (PROTAC®).
The PROTAC® of interest is incubated with CRBN and PARP1, bringing them in close proximity. PARP1 contains a GST tag, which is recognized by the GSH donor bead. CRBN contains a FLAG tag that binds to the AlphaLISA™ acceptor bead, which is conjugated with an anti-FLAG antibody. Upon excitation of the donor bead, a singlet oxygen is generated by the donor bead. The singlet oxygen excites the acceptor bead and emits light proportionally to the level of interaction

Need us to run inhibitor screens or profile your compounds against PROTAC® Optimization for PARP1-Cereblon Binding? Check out our PARP/PARPTrap™ Screening Services or PROTACs ® & Protein Degradation Services.

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Synonyms

ADP-ribosyltransferase diphtheria toxin-like 1, ARTD1, DNA ADP-ribosyltransferase PARP1, NAD(+) ADP-ribosyltransferase 1, ADPRT 1, Poly[ADP-ribose] synthase 1, Protein poly-ADP-ribosyltransferase PARP1, PPOL, PARP-1

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Product Data Gallery

Interaction of Cereblon with PARP1 via iRucaparib-AP6 (PROTAC®).

Inhibition by Rucaparib or Pomalidomide of iRucaparib-AP6 (PROTAC®)-mediated interaction of Cereblon with PARP1.

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Materials Required But Not Supplied

AlphaLISA™ anti-FLAG acceptor beads, 250 µg (PerkinElmer #AL112C)
Alpha™ GSH donor beads, 1mg (PerkinElmer #6765300)
Optiplate 384 (PerkinElmer #6007290)
AlphaScreen microplate reader
Adjustable micropipettor and sterile tips

Format

Catalog #	Name	Amount	Storage
100329	Cereblon/DDB1/Cul4A, Rbx 1 Complex*	20 µg	-80°C
80501	PARP1, GST-Tag*	20 µg	-80°C
	1 mM iRucaparib-AP6 PROTAC® (MW=887 Da)	15 µl	-80°C
	5X PP-02 Buffer	4 ml	-20°C
	1 mM Rucaparib (MW=323 Da)	15 µl	-20°C

*The concentration of the proteins is lot-specific and will be indicated on the tube.

UniProt #

P09874

Background

PARP1 (Poly (ADP-ribose) polymerase 1) acts as a first responder that detects DNA damage. PARP1 contributes to the efficiency of DNA repair by promoting the ADP-ribosylation of histones leading to the decompaction of chromatin structure, and by interacting with and modifying multiple DNA repair factors. PARP1 plays a role in cancer and is a well validated therapeutic target for cancer drugs.

Cereblon (CRBN) is the substrate-binding component of the E3 protein ligase complex DDB1-CUL4A-RBX1, which is involved in the ubiquitination and proteasomal degradation of target proteins. Binding of CRBN to a substrate engages its E3 ligase activity and results in the ubiquitination and ultimate degradation of the substrate. Many proteins are known targets of CRBN, including several transcription factors, growth factors and kinases. CRBN has become a target of choice for the development of many PROTACs of therapeutical interest.

Storage/Stability

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Applications

Identify and optimize PROTACs targeting PARP1.
Design novel molecules targeting CRBN.
Directly compare the activity of different PROTACs.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email [email protected]

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.

Warnings

Avoid freeze/thaw cycles

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

PARP

Regulatory Status

Research Use Only

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PROTAC® Optimization Kit for PARP1-Cereblon Binding

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