Bridge-It® S-Adenosyl Methionine (SAM) Fluorescence Assay Kit, 384-well format

SAM 1-1-1004B

Clear selection

Description

Bridge-It® S-Adenosyl Methionine (SAM) Fluorescence Assay Kit, 384-well format

Application:

  • Quantification of SAM level in biological fluids, cell culture and fermentation medium, and extracts of tissues and cells

  • Epigenetics and methylation studies

  • Numerous neurological and psychiatric disorders including Alzheimer’s disease, depression, HIV-related neurological dysfunction/dementia, multiple sclerosis, Parkinson’s disease, spinal cord degeneration, epilepsy, fibromyalgia, migraine headaches, and also chronic liver dysfunction, arteriosclerosis and cancer

 

Features:

  • Easy: Mix test sample or standard with the Assay solution and incubate at ~25°C

  • Fast: Read fluorescent signal after 30 minutes of incubation

  • Sensitive: Assay measures SAM at a lower limit of detection of 0.5 µM (i.e. 50 pmol / well in a 96-well or 20 pmol in a 384-well black microplate)

  • Flexible: Assay is adaptable to both low- and high-throughput screening formats

 


 

Background information on S-Adenosyl Methionine and the Bridge-It Fluorescence Assay

S-Adenosyl Methionine 
S-adenosyl methionine (also referred to as SAM, SAMe or AdoMet) plays a crucial role in the biological process of methylation in all types of organisms. In the methylation cycle, SAM serves as the donor of the methyl group used in the covalent modification of DNA and proteins. Variability in SAM levels have been linked to the processes of aging, numerous neurological and psychiatric disorders including Alzheimer’s disease, depression, HIV-related neurological dysfunction/dementia, multiple sclerosis, Parkinson’s disease, spinal cord degeneration, epilepsy, fibromyalgia, migraine headaches, and also chronic liver dysfunction, arteriosclerosis and cancer. Currently, SAM is quantified using the high pressure liquid chromatography (HPLC) method. HPLC is time consuming, costly and, due to the large amount of organic solvent required, not environmentally friendly.

 

 

Bridge-It® S-Adenosyl Methionine Fluorescence (SAM) Assay1,2
Mediomics Bridge-It® S-adenosyl methionine (SAM) fluorescence assay method is based on a combination of well-established fluorescence measurement techniques and our patented new assay platform design that utilizes DNA-binding proteins as biosensors for their respective small molecule co-regulators (ligands). The affinity of the DNA sequence-specific MetJ protein for its unique DNA-binding site is greatly increased in the presence of its ligand, S-adenosyl methionine. For this assay, the MetJ consensus sequence was split into two DNA “half-sites”. One half fragment was labeled with fluorescein and the other half fragment was labeled with Oyster® 645 fluorophore3,4. The relative amount of SAM present in a test sample will influence the amount of DNA-MetJ protein complex formation in the assay. When this complex forms, it brings the fluorescence labeled-DNA half-sites into close proximity and causes a measurable change (increase) in fluorescence signal emission that can be readily measured using a microplate reader (wavelength settings: absorption 485 nm; emission 665 nm). SAM concentrations in test samples are then determined using a SAM standard curve. The Bridge-It® SAM fluorescence assay method exhibits highly desirable performance characteristics including a high (>6:1) signal to background (S/B) ratio, a broad detection range (~0.4 µM – 100 µM), and, a detection sensitivity of 0.4 µM. This detection level (0.4 µM) is useful for quantifying SAM in most test samples of interest including biological fluids, cell culture and fermentation medium, and extracts of tissues and cells.

In contrast to the HPLC procedure, the Bridge-It® SAM fluorescence assay method utilizes the 384 and 96-well microplate format. Thus, it is ideally suited for the rapid, simultaneous measurement of SAM levels in large numbers of test samples. Also, this method is readily adaptable to the high-throughput screening platforms currently being used in drug discovery research.

 

Pricing: The 100-well kit is a one-time trial purchase. For bulk pricing options, please contact us. For any other questions or comments, please do not hesitate to contact us. We are always happy to help!

 

For information on the assay principle please see the Bridge-It Assay Technology platform page or download the protocol. This product is protected by patents and pending patents.


Selected Customer Publications using Mediomics’ Bridge-It SAM Fluorescence Assay Kit

1. Koczor CA, et al. AZT-induced Mitochondrial Toxicity: An Epigenetic Paradigm for Dysregulation of Gene Expression through Mitochondrial Oxidative Stress. Physiological Genomics. 2015 July 21.

— “For S-adenosylmethionine (SAM) 142 quantitation, a SAM fluorescence assay was utilized (Mediomics, St. Louis, MO).”

2. Dann SG, et al. Reciprocal regulation of amino acid import and epigenetic state through Lat1 and EZH2. The EMBO Journal. 2015 May 15.

3. Nishikawa K, et al. DNA methyltransferase 3a regulates osteoclast differentiation by coupling to an S-adenosylmethionine– producing metabolic pathway. Nature Medicine 2015 Feb 23; 21, 281–287.

— “ATP and SAM in osteoclast precursors and osteoclasts were detected using the Kinshiro ATP Luminescence kit (CosmoBio) and Bridge-It SAM Fluorescence Assay Kit (Mediomics), respectively.”

4. Mews P, Zee BM, Liu S, Donahue G, Garcia BA, Berger SL. Histone methylation has dynamics distinct from those of histone acetylation in cell cycle reentry from quiescence. Mol Cell Biol. 2014 Nov;34(21):3968-80.

–“To quantify SAM levels, the Bridge-It SAM fluorescence assay (Mediomics) was used according to the manufacturer’s instructions.”

5. Batra V, Verma P. Dietary L-methionine supplementation mitigates gamma-radiation induced global DNA hypomethylation: Enhanced metabolic flux towards S-adenosyl-l-methionine (SAM) biosynthesis increases genomic methylation potential. Food Chem Toxicol. 2014 Jul;69:46-54.

–“…l-methionine and SAM assay kit was purchased from Mediomics Inc. (St. Louis, MO, USA).”

6. Lo TF, Tsai WC, Chen ST. MicroRNA-21-3p, a berberine-induced miRNA, directly down-regulates human methionine adenosyltransferases 2A and 2B and inhibits hepatoma cell growth. PLoS One. 2013 Sep 30;8(9):e75628.

–“SAM levels in the supernatants were quantified using the Bridge-It SAM fluorescence assay kit (Mediomics) and detected using SpectraMax plate reader (Molecular Devices) according to the manufacturer’s instructions.”

7. Rascan et al. S-Adenosylmethionine: A Novel Factor in the Individualization of Thiopurine Therapy.

–“The commercially available assay for SAM determination is a Mediomics Bridge-It® fluorescence assay based on a combination of fluorescence measurement techniques…”

8. Schrötter A, Pfeiffer K, El Magraoui F, Platta HW, Erdmann R, Meyer HE, Egensperger R, Marcus K, Müller T. The amyloid precursor protein (APP) family members are key players in S-adenosylmethionine formation by MAT2A and modify BACE1 and PSEN1 gene expression-relevance for Alzheimer’s disease. Mol Cell Proteomics. 2012 Nov;11(11):1274-88.

–“S-adenosylmethionine (SAM) levels were measured with Bridge-It™ S-adenosylmethionine Fluorescence Assay (Mediomics, St. Louis, MO, USA) using a 384-well microplate format.”

9. Kazutaka S, Winnall WR, Muir JA, Hedger MP. Regulation of Sertoli cell activin A and inhibin B by tumour necrosis factor α and interleukin 1α: interaction with follicle-stimulating hormone/adenosine 3′,5′-cyclic phosphate signalling. Mol Cell Endocrinol. 2011 Mar 30;335(2):195-203.

–“The commercially available assay for SAM determination is a Mediomics Bridge-It® fluorescence assay based on a combination of fluorescence measurement techniques.”

10. Batra V, Sridhar S, Devasagayam TP. Enhanced one-carbon flux towards DNA methylation: Effect of dietary methyl supplements against γ-radiation-induced epigenetic modifications. Chem Biol Interact. 2010 Feb 12;183(3):425-33.

–“…SAM assay kit was purchased from Mediomics Inc. (St. Louis, MO, USA)…”

11. Watson JA, Watson CJ, McCrohan AM, Woodfine K, Tosetto M, McDaid J, Gallagher E, Betts D, Baugh J, O’Sullivan J, Murrell A, Watson RW, McCann A. Generation of an epigenetic signature by chronic hypoxia in prostate cells. Hum Mol Genet. 2009 Oct 1;18(19):3594-604.

–“SAM levels were quantified using the Bridge-It® SAM fluorescence assay (Mediomics).”

12. Steffens AA. Low-dose of sodium arsenite causes delayed differentiation in C2C12 mouse myoblast cells through the repression of the transcription factor myogenin. 2009. Thesis. Clemson University.

–“S-adenosyl methionine (SAM) concentrations were determined using the Bridge-It SAM Fluorescence Assay (Mediomics, St. Louis, MO).”

13. Desaulniers D, Xiao GH, Lian H, Feng YL, Zhu J, Nakai J, Bowers WJ. Effects of mixtures of polychlorinated biphenyls, methylmercury, and organochlorine pesticides on hepatic DNA methylation in prepubertal female Sprague-Dawley rats. Int J Toxicol. 2009 Jul-Aug;28(4):294-307.

14. Jean-François Coppin, Wei Qu, and Michael P. Waalkes. Interplay between Cellular Methyl Metabolism and Adaptive Efflux during Oncogenic Transformation from Chronic Arsenic Exposure in Human Cells. J Biol Chem. 2008 July 11; 283(28): 19342–19350.

–“SAM levels were assayed according to manufacturer instructions using the Bridge-it® SAM fluorescence assay kit (Mediomics, St. Louis, MO).”

15. Chan A, Tchantchou F, Graves V, Rozen R, Shea TB. Dietary and genetic compromise in folate availability reduces acetylcholine, cognitive performance and increases aggression: critical role of S-adenosyl methionine. J Nutr Health Aging. 2008 Apr;12(4):252-61.

–“SAM levels in homogenates of forebrain were determined by comparison to SAM standards using a Bridge-It S-Adenosyl Methionine (SAM) Fluorescence Assay Kit (Mediomics, LLC).”

16. Chan A, Shea TB. Supplementation with apple juice attenuates presenilin-1 overexpression during dietary and genetically-induced oxidative stress. J Alzheimers Dis. 2006 Dec;10(4):353-8.

–“… SAM levels in homogenates of forebrain and in AJC were determined by comparison to SAM standards us- ing a Bridge-It S-Adenosyl Methionine (SAM) Fluo- rescence Assay Kit (Mediomics, LLC) assayed on a Hitachi F2000 fluorescent spectrophotometer.”


  1. Bridge- It® is a registered trademark of Mediomics, LLC, St. Louis, Missouri, U.S.A. Mediomics has a worldwide, exclusive license for this assay platform from Saint Louis University, St. Louis, Missouri.

  2. The Bridge-It® S-adenosyl methionine (SAM) fluorescence assay kit is provided for laboratory R&D use only. This product is not approved by the U.S. Government or by the government of any other country of the world for use in clinical diagnosis of disease or treatment of disease in humans or animals.

  3. Oyster® is a registered trademark of Denovo Biolabels, GmbH, Munster, Germany.

  4. Flownamics® Analytical Instruments, Inc., Madison, Wisconsin, is the authorized U.S. distributor of Oyster® dyes.

Catalog #: 1-1-1004A (100 measurements) , 1-1-1004B (384 measurements)

Protocol :

SDS: 

Supporting Protocol :