JMJD2B (KDM4B) Homogenous Assay Kit

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Catalog #

50414

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Description

The JMJD2B (KDM4B) Homogeneous Assay Kit is designed to measure JMJD2B (Jumonji domain-containing protein 2B) activity for screening and profiling applications. The JMJD2B (KDM4B) Homogeneous Assay Kit comes in a convenient 96-well or 384-well AlphaLISA® format, with enough purified recombinant JMJD2B (amino acids 2-500), substrate, primary antibody, assay and detection buffer for 100 or 400 enzyme reactions.

50414 scheme 1

Figure 1: JMJD2B (KDM4B) Homogenous Assay Kit schematic.
A sample containing JMJD2B is incubated with a substrate. This is followed by the addition of acceptor beads and primary antibody, and finally donor beads. Alpha-counts are then counted. Alpha-counts are proportional to JMJD2B demethylase activity.

Need us to run inhibitor screens or profile your compounds against JMJD2B (KDM4B)? Check out our Demethylase Screening Services.

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Synonyms

JHDM3B, KDM4B

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Product Data Gallery

Inhibition of JMJD2B activity by 2, 4-Pyridine Dicarboxylic Acid (96 reactions)

Inhibition of JMJD2B activity by 2, 4-Pyridine Dicarboxylic Acid (384 reactions)

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Materials Required But Not Supplied

AlphaLISA® anti-mouse IgG acceptor beads, 5 mg/ml (PerkinElmer #AL105C)
AlphaScreen® Streptavidin-conjugated donor beads, 5 mg/ml (PerkinElmer #6760002S)
Optiplate -96 or -384 (PerkinElmer #6005290)
AlphaScreen^® microplate reader

Format

96 reactions

Catalog #	Name	Amount	Storage
50111	JMJD2B (KDM4B), GST-Tag*	10 µg	-80°C
52140E	Primary Antibody 5	5 µl	-80°C
79841	Biotinylated Histone H3 Peptide Substrate	300 reactions	-80°C
52407	4x HDM Assay Buffer 2	3 x 1 ml	-80°C
52301	4x Detection Buffer	2 ml	-20°C

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

384 reactions

Catalog #	Name	Amount	Storage
50111	JMJD2B (KDM4B), GST-Tag*	60 µg	-80°C
52140E	Primary Antibody 5	20 µl	-80°C
79841	Biotinylated Histone H3 Peptide Substrate	500 reactions	-80°C
52407	4x HDM Assay Buffer 2	3 x 1 ml	-80°C
52301	4x Detection Buffer	2 ml	-20°C

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

UniProt #

O94953

Background

JMJD2B (Jumonji domain -containing protein 2B), also known as KDM4B, belongs to the JMJD2 subfamily of JMJD histone lysine demethylase proteins. It is involved in demethylating H3K9me2/3, thus participating in osteogenic differentiation of mesenchymal stem cells, regulation of transcription for maintenance of trophoblast stem cells, DNA damage response (DDR), cell cycle progression and apoptosis. JMJD2B is found at high levels in different cancer types, such as breast, gastric and colorectal cancer and Hodgin’s lymphoma, and also during inflammation. JMJD2B is regulated by HIF1α (hypoxia-inducible factor 1 alpha), and silencing JMJD2B in colorectal cancer cells can stimulate DDR responses and inhibit cancer cell proliferation. The use of JMJD2B inhibitors is an area of interest that requires further development, and the development of active inhibitors will be beneficial in the cancer therapy field.

References

Manni W., et al., 2022 Signal Transduct Target Ther 7:304.
Mak K., et al., 2021 Scientific Reports 11:884.
Chen L., et al., 2014 British Journal of Cancer 110:1014-1026.

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

Study enzyme kinetics and screen small molecule inhibitors in high throughput screening (HTS) applications.

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 JMJD2B Homogenous Assay Kit is compatible with up to 1% final DMSO concentration.
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

Demethylase

Regulatory Status

Research Use Only