SMYD3 Homogeneous Assay Kit

Catalog #

52063

$1,125 *

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 SMYD3 Homogeneous Assay Kit is designed to measure SMYD3 (SET and MYND domain containing protein 3) activity for screening and profiling applications. The SMYD3 Homogeneous Assay Kit comes in a convenient 384-well AlphaLISA® format, with enough purified recombinant SMYD3/HSP90 complex, GST-tagged SMYD3 substrate, primary antibody and methylation assay buffer for 384 enzyme reactions.

Figure 1: SMYD3 Homogenous Assay Kit schematic.
A sample containing SMYD3/HSP90 complex is incubated with a substrate for four hours. 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 SMYD3 methylase activity.

Need us to run inhibitor screens or profile your compounds against SMYD3? Check out our Methyltransferase Screening Services.

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Synonyms

Histone-lysine N-methyltransferase SMYD3, KMT3E, SET and MYND domain-containing protein 3, Zinc finger MYND domain-containing protein 1, ZMYND1, ZNFN3A1

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Product Data Gallery

SMYD3 activity

SMYD3 activity is inhibited by MEKK2 Mutant

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Materials Required But Not Supplied

AlphaLISA® anti-rabbit IgG acceptor beads (Perkin Elmer #AL104C)
AlphaScreen® Glutathione-conjugated donor beads (Perkin Elmer #6765300)
Optiplate-384 (PerkinElmer #6007290)
AlphaScreen® microplate reader

Format

Catalog #	Component	Amount	Storage
51110	SMYD3/HSP90*	2 x 50 µg	-80°C
52120	100 µM S-adenosylmethionine	2 x 250 µl	-80°C
52140O3	Primary antibody 15-3	20 µl	-80°C
	SMYD3 substrate	2 x 200 µl	-80°C
	4x SMYD3 Assay Buffer	3 x 1 ml	-20°C
	4x Detection Buffer 3D	2 ml	-20°C
	0.5 mM DTT	200 µl	-20°C
	Plate sealer		Room Temp

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

UniProt #

Q9H7B4

Background

SMYD3, also known as SET (suppressor of variegation, enhancer of Zeste, Thithorax) and MYND (myeloid-Nervy-DEAF-1) domain-containing protein 3 is a lysine methyltransferase family member. It is involved in methylation of H3K4 and H4K5, but also of non-histone substrates such as VEGFR1 (vascular endothelial growth factor receptor 1), MAP3K2 (MAP3 kinase 2), AKT1 (protein kinase B) and HER2 (human epidermal growth factor receptor 2). In adults it is found in platelets, testis, CD8⁺ T-cells, and at very low levels in the heart, bladder, prostate, spinal cord and retina. It is overexpressed in several cancer types, such as liver and breast cancer, and plays a role in disease progression via its substrates. For instance, SMYD3 activates AKT1 by methylation on lysine 14, resulting in cell growth, survival and neovascularization. The low or absence expression of SMYD3 in normal tissues makes it a promising cancer therapy candidate. Several inhibitors are undergoing clinical trials, but further studies are necessary. A deep understanding of the role of SMYD3 and identification of potent specific inhibitors will advance the cancer therapy field.

References

Dillon S.C., et al. 2005 Genome Biology 6:227.
Silva F.P., et al. 2008 Oncogene 27:2686.
Bernard B., et al., 2021 Clin Epigenetics 13:45.

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 SMYD3 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

Methyltransferase

Regulatory Status

Research Use Only