VHL Binding Assay Kit

Catalog #

78805

$1,495 *

Size: 96 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 Von Hippel–Lindau tumor suppressor (VHL) Binding Assay Kit is a homogeneous assay which measures the binding of VHL to the high-affinity VHL fluorescent probe (BDY FL VH032) using Fluorescence Polarization (FP). In addition, the kit includes the VHL inhibitor VH298 for use as an inhibitor control. This assay takes advantage of the fluorescent substrate BDY FL VH032. Using this kit, only one step on a 96-well plate is required. The BDY FL VH032 is incubated with the VHL complex in the presence of a compound of interest to produce a change in fluorescent polarization. The FP signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.

Need us to run inhibitor screens or profile your compounds against VHL Binding? Check out our Ubiquitination Screening Services.

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Synonyms

von Hippel-Lindau disease tumor suppressor, Protein G7, pVHL

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Product Data Gallery

Inhibition of VHL binding to fluorescent probe BDY FL VH032 by VH298, measured using the VHL Binding Assay kit (BPS Bioscience #78805).

Product Info

Storage and Usage

Citations

Assay Kit Format

Fluorescence Polarization

Supplied As

The kit provides enough purified ELOB/ELOC/VHL complex, VHL fluorescent probe (BDY FL VH032), and assay buffer for 100 enzyme reactions.

Materials Required But Not Supplied

Adjustable micropipettor and sterile tips

Format

Catalog #	Name	Amount	Storage
100361	ELOB/ELOC/VHL Complex*	150 µg	-80°C
	BDY FL VH032, 250 µM	10 µl	-80°C
	VHL Assay Buffer (FP)	10 µl	-20°C
	VH298, 50 mM	15 µl	-20°C
79685	Black, low binding, microtiter plate	1	Room Temp.

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

UniProt #

P40337

Background

VHL is a tumor suppressor that functions as the substrate recognition subunit of the Cullin2 RING E3 ligase complex (CRL2^VHL). CRL2^VHL plays important roles in oxygen sensing by targeting hypoxia-inducible factor-alpha (HIF-α) subunits for ubiquitination and degradation. Elongin B (ELOB) and Elongin C (ELOC) are heterodimers that bind to the BC-box motif present in VHL. In this specific E3 ligase complex, the ELOB/ELOC works as an adapter between target proteins and Cullin-2/Rbx1 in VHL-box E3 ubiquitin ligases.

VHL is targeted by bifunctional molecules such as proteolysis-targeting chimeras to induce degradation of proteins of interest. Design and characterization of VHL binders is an important step in the design of VHL-targeting molecular degraders.

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

Titrate or screen small molecule inhibitors of VHL binding to the probe in drug discovery and high throughput (HTS) applications.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit Assay Kits FAQ (bpsbioscience.com) for detailed troubleshooting instructions and information about FP assays. Read our eBook "Fluorescence Polarization Assays" to learn more about principles of FP.

For further questions, please email [email protected].

Contraindications

The final concentration of DMSO in the assay should not exceed 1%.

Warnings

This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note: FP, assay principles and applications.  

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

Ubiquitination

Regulatory Status

Research Use Only

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VHL Binding Assay Kit

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