PROTAC® Optimization Kit for BET Bromodomain-Von Hippel Lindau (VHL) Binding

Catalog #

79790

$860 *

Size: 384 reactions

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Description

The PROTAC® Optimization Kit for BET Bromodomain-Von Hippel Lindau (VHL) Binding is designed for testing and profiling PROTACs® directed against the BET Bromodomain family and VHL complex. VHL is a substrate recognition component of E3 protein ligase complex and is linked by Elongin C to a heterodimeric Cul2/Rbx1 module, which functions as a potent activator of the ubiquitination of target proteins by an E2 conjugating enzyme. Elongin B interacts with the complex through Elongin C and stabilizes the binding of Elongin C to VHL. Mutations in VHL are associated with the inherited von Hippel-Lindau cancer syndrome and numerous forms of renal cell carcinoma.

With this kit, only three simple steps on a microtiter plate are required for PROTAC® activity detection. First, a sample containing PROTAC® is incubated with VHL and BRD3(BD2), one of the BET bromodomains. Next, acceptor beads are added, then donor beads, followed by reading the Alpha-counts.

PROTAC Optimization Kit for BET Bromodomain-Von Hippel Lindau (VHL) Binding

Need us to run inhibitor screens or profile your compounds against PROTAC® Optimization for BET Bromodomain-Von Hippel Lindau (VHL) Binding? Check out our Bromodomain Screening Services or PROTACs ® & Protein Degradation Services.

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Synonyms

Bromodomain containing 3, BRD3, RING3L

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Supplied As

The PROTAC® Optimization Kit for BET Bromodomain-VHL Binding comes in a convenient AlphaLISA® format, with ARV-771 PROTAC®, purified BRD3(BD2) and VHL proteins, and buffer for 384 reactions. (+)JQ1 is included as a control inhibitor of PROTAC® binding to BRD3(BD2).

Materials Required But Not Supplied

AlphaLISA anti-FLAG acceptor beads, 5 mg/ml (PerkinElmer #AL112C)
Alpha Glutathione donor beads, 5 mg/ml (PerkinElmer #6765300)
Optiplate 384 (PerkinElmer #6007290)
AlphaScreen microplate reader
Adjustable micropipettor and sterile tips

Format

Catalog #	Component	Amount	Storage
100373	VHL Complex, FLAG-tag	25 µg	-80°C	Avoid freeze/ thaw cycles
31033	BRD3(BD2), GST-tag	10 µg	-80°C
	ARV-771 (MW = 987 Da)	10 µg	-80°C
79311	3x Immuno Buffer 1	4 ml	-20°C
27403	(+)-JQ1 (10 mM)	20 µg	-20°C

*The initial concentrations of VHL and BRD3 are lot specific and will be indicated on the tubes containing the proteins

References

Fisher, S.L., Phillips, A.J. Curr Opin Chem Biol 2018; 44:47-55

Storage/Stability

At least six months from date of receipt when stored as directed.

Instructions for Use

See assay protocol for detailed instructions.

Shipping Temperature

-80°C

Contraindications

Green and blue dyes that absorb light in the AlphaScreen signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of culture medium RPMI 1640 at >1% leads to signal reduction due to the presence of excess biotin and iron in this medium. 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

PROTACs

Regulatory Status

Research Use Only

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PROTAC® Optimization Kit for BET Bromodomain-Von Hippel Lindau (VHL) Binding

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