Methyltransferase Fluorometric Assay Kit

Cayman Chemical Item Number 700150

MT Fluorometric Assay Kit

Methyltransferase Fluorometric Assay Kit

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Description

Methylation of key biological molecules and proteins play important roles in numerous biological systems, including signal transduction, biosynthesis, protein repair, gene silencing, and chromatin regulation.1 The S-adenosylmethionine (SAM) dependent methyltransferases use SAM, the second most commonly used enzymatic cofactor after ATP. SAM, also known as AdoMet, acts as a donor of a methyl group that is required for the modification of proteins and DNA.2 Aberrant levels of SAM have been linked to many abnormalities, including Alzheimer’s Disease, depression, Parkinson’s Disease, multiple sclerosis, liver failure, and cancer.1,2 Cayman’s Methyltransferase Fluorometric Assay Kit is a continuous enzyme-coupled assay that can continuously monitor SAM-dependent methyltransferases.3 The removal of the methyl group from SAM generates S-adenosylhomocysteine (AdoHcy), which is rapidly converted to S-ribosylhomocysteine and adenine by AdoHcy nucleosidase. This rapid conversion prevents the buildup of AdoHcy and its feedback inhibition on the methylation reaction. Finally, the adenine is converted to hypoxanthine, by adenine deaminase, which in turn is converted to urate and hydrogen peroxide (H2O2). The reaction between H2O2 and ADHP (10-acetyl-3,7,-dihydroxyphenoxazine) produces the highly fluorescent compound resorufin. Resorufin fluorescence can be easily analyzed with an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm. The assay is supplied with AdoHcy as a positive control. The assay can be used with any purified SAM-dependent methyltransferase.

1 Loenen, W.A.M. S-Adenosylmethionine: Jack of all trades and master of everything? Biochem Soc Trans 34(2) 330-333 (2006).

2 Chiang, P.K., Gordon, R.K., Tal, J., et al. S-adenosylmethionine and methylation. FASEB J 10 471-480 (1996).

3 Dorgan, K.M., Wooderchak, W.L., Wynn, D.P., et al. An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases. Anal Biochem 350 249-255 (2006).

Synonyms
  • MT Fluorometric Assay Kit
Formal Name
Stability 6 months
Storage -80°C
Shipping Dry ice in continental US; may vary elsewhere

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Background Reading

Couture, J., Collazo, E., Hauk, G., et al. Structural basis for the methylation site specificity of SET7/9. Nat Struct Mol Biol 13(2) 140-146 (2006).

Dorgan, K.M., Wooderchak, W.L., Wynn, D.P., et al. An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases. Anal Biochem 350 249-255 (2006).

Chiang, P.K., Gordon, R.K., Tal, J., et al. S-adenosylmethionine and methylation. FASEB J 10 471-480 (1996).

Loenen, W.A.M. S-Adenosylmethionine: Jack of all trades and master of everything? Biochem Soc Trans 34(2) 330-333 (2006).

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700150-1ea
96-Well Cover Sheet
Half Volume 96-Well Solid Plate (black)
DMSO Assay Reagent 1 mL
HCl Assay Reagent (20 mM) 1 mL
Resorufin Standard 500 µL
Methyltransferase Assay Buffer 1 ea
Methyltransferase Assay Buffer Additive 1 ea
Methyltransferase Enzyme Mixture 3 × 1 ea
Methyltransferase Assay AdoHcy Positive Control 1 ea
Methyltransferase Assay S-Adenosylmethionine 3 × 1 ea
Methyltransferase Fluorometric Mixture 3 × 1 ea
Methyltransferase Resorufin Buffer (10X) 1 ea
Size Price Quantity Subtotal
96 wells $391.00 $0.00
Bulk Contact
Cart Total $0.00

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Pricing updated 2014-04-23. Prices are subject to change without notice.

To ask for assistance with one of our products please contact a Technical Support Scientist.

Warning This product is not for human or veterinary use.

You may be eligible to receive a free sample Methyltransferase Fluorometric Assay Kit under the Cayman Challenge program.

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Cayman Chemical Company regularly evaluates our shipping methods to ensure product stability is maintained and our corporate environmental impact is minimized. Products selected for review are evaluated and shipping methods are certified by our Scientists. While occasionally our method of shipping a product may change, rest assured the shipping conditions on all Cayman Chemical products have been reviewed and validated.

Upon receipt, it is important to check the storage requirements of your products and place them at the appropriate temperature. Products shipped without ice may require frozen storage for long term stability.

Cayman Chemical was formed over thirty years ago by demonstrating the value of naturally growing gorgonian corals as a renewable, economically viable source of prostaglandins. Today we operate our business knowing that we owe our start to the renewable resources our environment had provided. As Cayman Chemical continues to grow we consistently evaluate our business processes to ensure we are operating at the highest environmental standards.

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