BCL-2 TR-FRET Assay Kit

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Catalog #

50222

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Description

The BCL-2 TR-FRET Assay Kit is designed to measure the inhibition of BCL-2 (B-cell lymphoma 2) binding to its ligand in a homogeneous 96 or 384 reaction format. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. This kit contains enough recombinant BCL-2, BCL-2 peptide ligand, assay buffer, Anti-His Terbium-Labeled Donor, and Dye-labeled Streptavidin Acceptor for 96 or 384 reactions

Figure 1. Illustration of the BCL-2 TR-FRET Assay Kit principle.
A sample containing terbium-labeled donor, dye-labeled acceptor, BCL-2, peptide ligand, and an inhibitor is incubated for 180 minutes. The Donor Terbium-labeled anti-His antibody binds to His-tagged BCL-2. The BCL-2 peptide ligand is labeled with biotin, which allows the dye-labeled streptavidin acceptor to bind to the BCL-2 peptide ligand. The TR-FRET signal is generated by proximity induced upon interaction of BCL-2 with the BCL-2 peptide ligand. Then, the fluorescence intensity is measured using a fluorescence reader capable of TR-FRET measurements. The signal generated is proportional to BCL-2 activity.

Need us to run inhibitor screens or profile your compounds against BCL-2? Check out our Apoptosis Screening Services.

This product has been cited 4 times.

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Synonyms

Bcl, Bcl2, B cell lymphoma 2, HTRF, BCLXL, BCL2

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Product Data Gallery

Inhibition of BCL-2 binding to its peptide ligand by ABT-199 (96 reactions)

Inhibition of BCL-2 binding to its peptide ligand by ABT-199 (384 reactions)

Product Info

Storage and Usage

Citations4

Assay Kit Format

TR-FRET

Materials Required But Not Supplied

Fluorescent microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
Adjustable micropipettor and sterile tips

Format

96 Reactions

Catalog #	Component	Amount	Storage
50272	BCL-2, His-tag*	1 µg	-80°C
30057	BCL-2 Peptide Ligand, Biotin-Labeled	25 µl	-80°C
30017	Anti-His Terbium-Labeled donor	10 µl	-20°C
	Dye-labeled Streptavidin Acceptor	10 µl	-20°C
30059	BCL TR-FRET Assay Buffer	4 ml	-20°C
79696	White, nonbinding, low volume microtiter plate	1	Room Temp

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

384 Reactions

Catalog #	Component	Amount	Storage
50272	BCL-2, His-tag*	2 µg	-80°C
30057	BCL-2 Peptide Ligand, Biotin-Labeled	50 µl	-80°C
30017	Anti-His Terbium-labeled donor	2 x 10 µl	-20°C
	Dye-labeled Streptavidin Acceptor	2 x 10 µl	-20°C
30059	BCL TR-FRET Assay Buffer	4 ml	-20°C
79969	White, nonbinding, low volume microtiter plate	1	Room Temp

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

UniProt #

P10415

Background

B-cell lymphoma 2 (BCL-2) is a member of the BCL family that regulates apoptosis through pro-apoptotic or anti-apoptotic signals. BCL-2 is normally localized at the outer membrane of mitochondria and blocks pro-apoptotic signals, promoting cell survival. Genetic alteration of BCL-2 has been observed in various types of cancer, including lymphoma, leukemia, melanoma, as well as breast and prostate cancer. The protein is also involved in cancer cell resistance to treatment due to its pro-survival functions. ABT-199, also known as Venetoclax, is a BH3-mimetic that specifically blocks the function of BCL-2 and was the first BCL-2 targeted agent to receive FDA approval (in 2016). It is now approved for the treatment of chronic lymphocytic leukemia and small lymphocytic lymphoma

References

Filippakopoulos, P., et al.,Cell 2012; 149:214.

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

Screen small molecule inhibitors in drug discovery and high-throughput screening (HTS) applications.

Shipping Temperature

-80°C

Notes

Since January of 2018, this kit has been optimized for better performance. The previous version of the BCL-2 TR-FRET Assay Kit, Cat. #50222, can still be purchased upon special request.

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

Apoptosis

Regulatory Status

Research Use Only