SetDB1 Chemiluminescent Assay Kit

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Catalog #

51056

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Description

The SetDB1 Chemiluminescent Assay Kit is designed to measure SetDB1 activity for screening and profiling applications. The SetDB1 Chemiluminescent Assay Kit comes in a convenient 96-well or 384-well format, precoated with histone H3 peptide substrate, an antibody against the methylated lysine 9 residue of Histone H3, a secondary HRP-labeled antibody, S-adenosylmethionine, methyltransferase assay buffer, and purified SetDB1 enzyme (amino acids 510-1290) for 96 or 384 enzyme reactions. The advantage of the SetDB1 Chemiluminescent Assay Kit is a highly specific antibody that recognizes the methylated K9 residue of Histone H3. With this kit, only three simple steps are required. First, S-adenosylmethionine is incubated with a sample containing the assay buffer and the methyltransferase enzyme. Next, the primary antibody is added. Finally, the wells are treated with an HRP-labeled secondary antibody followed by addition of the HRP substrate to produce chemiluminescence that can be measured using a chemiluminescence reader

This product has been cited 1 time.

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Synonyms

Histone-lysine Nmethyltransferase SETDB1, SET domain bifurcated 1, KMT1E, ERG-associated protein with SET domain (ESET)

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Product Data Gallery

SetDB1 enzymatic activity. (96 reactions)

SetDB1 enzymatic activity. (384 reactions)

Product Info

Storage and Usage

Citations1

Assay Kit Format

Chemiluminescent

Materials Required But Not Supplied

TBST buffer (1 x TBS, pH 8.0, containing 0.05% Tween-20)
Luminometer
Orbital Shaker

Format

96 Reactions

Catalog #	Component	Amount	Storage
51055	SetDB1 Human Enzyme*	2 μg	-80°C
52120	20 μM S-adenosylmethionine	250 μl	-80°C
52140A	Primary Antibody 1	100 μl	-80°C
52130H	Secondary HRP-labeled Antibody 1	10 μl	-80°C
52160	4x HMT assay buffer 1	3 ml	-20°C
52100	Blocking Buffer 4	50 ml	+4°C
	0.5 M DTT	200 µl	-20°C
	HRP chemiluminescent substrate A (transparent bottle)	6 ml	+4°C
	HRP chemiluminescent substrate B (brown bottle)	6 ml	+4°C
	96-well plate precoated with histone substrate		+4°C

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

384 Reactions

Catalog #	Component	Amount	Storage
51055	SetDB1 Human Enzyme*	4 μg	-80°C
52120	20 μM S-adenosylmethionine	2 x 250 μl	-80°C
52140A	Primary Antibody 1	2 x 100 μl	-80°C
52130H	Secondary HRP-labeled Antibody 1	2 x 10 μl	-80°C
52160	4x HMT assay buffer 1	2 x 3 ml	-20°C
52100	Blocking Buffer 4	2 x 50 ml	+4°C
	0.5 M DTT	200 µl	-20°C
	HRP chemiluminescent substrate A (transparent bottle)	2 x 6 ml	+4°C
	HRP chemiluminescent substrate B (brown bottle)	2 x 6 ml	+4°C
	384-well plate precoated with histone substrate		+4°C

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

UniProt #

Q15047

Background

SetDB1 (Histone-lysine N-methyltransferase SETDB1) is a methyltransferase that reversibly catalyzes the di and trimethylation of histone 3 (H3) resulting in inhibition of gene transcription within the euchromatic regions of chromosomes. Studies linking the activity of SetDB1 and H3 methylation can elucidate the mechanisms of tumorigenesis in various types of cancers.

References

Torrano J., et al., 2019 Clinical Epigenetics. 11(1):43.
Dillon S.C., et al., 2005 Genome Biol. 6(8):227.

Storage/Stability

This assay kit will perform optimally for up to 6 months from date of receipt when materials are stored as directed. Avoid multiple freeze/thaw cycles.

Applications

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

Shipping Temperature

-80°C

Notes

License Disclosure
Visit bpsbioscience.com/license for the label license and other key information about this product.

Troubleshooting Guide
Visit bpsbioscience.com/cell-line-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Contraindications

This assay is compatible with DMSO >1%.
This assay is incompatible with strong acids or bases, ionic detergents and high salt.

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

Methyltransferases

Regulatory Status

Research Use Only