DOCSLIB.ORG

Ethidium Bromide Alternatives Assessment August 2009 (Revised: August 2011)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Ethidium Bromide Alternatives Assessment August 2009 (Revised: August 2011) MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity Ethidium dsDNA 290 nm agarose 0.2ng-0.5ng May be UV Mutagenic with Managed Approx. $30 Bromide 1 ssDNA 605 nm stored at Transilluminator S9 activation as for 10mL of RNA acrylamide room of Salmonella hazardous 10mg/mL temperature; Polaroid 667 TA98 and waste solution; use PCR indefinite black & white TA1537 0.5µg/mL for storage strains agarose gel; yields 5,000 LD 50 , rat (oral): 40-mL gels, 1503 mg/kg $0.006/gel. (slightly toxic) Aquatic toxicity : LC 50 not available, MSDS indicates “may cause long- term adverse effects on aquatic environment” SYBR Safe 2 dsDNA 280/502 nm agarose Comparable Keep away UV Weakly Approved $53.75 for ssDNA 530 nm to ethidium from heat Transilluminator mutagenic with by MWRA 10,000X acrylamide bromide and light; S9 activation for drain SYBR Safe stable for blue (vis) light of Salmonella disposal, in DMSO, pre-cast approx. 6 transilluminator TA98 and May 2005 400µL; 4µL months (SafeImager™ TA1537 for 40 mL post-gel when stored recommended) strains gel = at room $0.5375 per temperature. laser scanner LD 50 , rat (oral): gel. >5,000 mg/kg Polaroid 667 (practically black & white nontoxic) 1 Available from numerous vendors 2 Invitrogen/Molecular Probes (now Life Technologies) QUESTIONS??? Send comments to [email protected] 1 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity with SYBR Safe filter (S37100) or Aquatic Molecular toxicity : Probes SYPRO LC 50 >750mg/L filter (S6656) or Kodak Wratten #9 filter GelRed 3 dsDNA 275 nm agarose 1-3ug/mL; Store in UV Weakly Approved Available as ssDNA 600 nm visualizes water at Transilluminator mutagenic at by MWRA 0.5mL RNA acrylamide DNA doses room 18.5 µg/mL for drain (500µL) of as low as temperature similar spectra with S9 disposal, 10,000X pre-cast 25ng. or at 4 oC as ethidium activation of April 2010 concentrate; (agarose) bromide, Salmonella working therefore can TA98 dilution is 1- post-gel use same 3µg/mL for (both) visualization LD 50 , rat (oral): 167-500 settings not provided minigels on MSDS $100-$105 Cell ($0.20- membrane $0.63/gel) staining: did not cross HeLa cell membranes after 30 minutes incubation Aquatic toxicity : LC 50 >750mg/L GelGreen 3 dsDNA 250- agarose 1-3µg/mL Store in UV Non- Approved 500µL ssDNA 300/500nm water at Transilluminator mutagenic by MWRA 10,000X RNA ~525 nm acrylamide room (254 nm) from 0.4µg/mL for drain concentrate 3 Biotium QUESTIONS??? Send comments to [email protected] 2 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity – not temperature to 18.5 µg/mL disposal, in DMSO (GelRed recommended or at 4 oC 488nm laser- in Salmonella April 2010 recommended due to slow based gel TA98 and 1-3µg/mL for for ssDNA and diffusion of scanner TA1537 167-500 RNA) dye strains, w/ and minigels w/o S9 pre-cast activation. $90-$100 ($0.18- post-gel LD 50 , rat (oral): $0.60/gel); not provided on MSDS Cell membrane staining: did not cross HeLa cell membranes after 30 minutes incubation Aquatic toxicity : LC 50 >750mg/L MegaFluor 4 dsDNA Similar agarose Can run at Store 10X UV Manufacturer Not $225 for absorption 3pg-5pg gel at 4 oC transilluminator claims present in 500-1500 -added directly to and post-gel (picograms) molecule is the gel gels = DNA sample, not emission (agarose) similar spectra large and does running $0.15-$0.45 gel or buffer spectra as Can as ethidium not cross cell buffer; per gel ethidium visualize bromide, membrane. buffer -not advised for bromide DNA doses therefore can Indicated may be Dose : denatured DNA less than use same testing disposed MegaFluor 10ng if visualization performed on by drain. to DNA, 1:10 samples are settings epithelial cells 4 GenTaur QUESTIONS??? Send comments to [email protected] 3 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity first showed no No formal MegaFluor incubated at mutagencity. MWRA to DNA 60 oC. approval; marker, 2:10 LD 50 , rat( oral) : meets or not stated exceeds MegaFluor criteria for to genomic Aquatic SYBR DNA, 4:10 toxicity : Safe. LC 50 not stated 1.5 mL = 1,500 dsDNA gels @10µL (100 samples per 1.5mL kit) SafeWhite 5 dsDNA 320 nm agarose 0.1ng-0.3ng Store 6X in Same as for Non- Approved 6X in water; ssDNA 480 nm water at 4 oC ethidium bromide mutagenic in by MWRA 1mL, $58 RNA Salmonella for drain TA98 and disposal, Dose : 5µL TA1537 April 2010 per 100 mL strains, w/ and agarose = w/o S9 $0.29 per activation. gel LD 50 , rat( oral) : not stated Aquatic toxicity : LC 50 not stated SafeRed 4 dsDNA 490 nm agarose 0.3ng-0.8ng Store 6X in Same as for Non- Approved 6X in water; ssDNA 630 nm water at 4 oC ethidium bromide mutagenic in by MWRA 1 mL, $58 RNA Salmonella for drain TA98 and disposal, Dose : 5µL TA1537 April 2010 per 100 mL 5 Applied Biological Materials QUESTIONS??? Send comments to [email protected] 4 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity strains, w/ and agarose = w/o S9 $0.29 per activation. gel LD 50 , mouse subcutaneous : 16mg/kg Aquatic toxicity : LC 50 not stated SafeGreen 4 dsDNA 290 nm agarose 0.2ng-0.6ng Store 6X in Same as for Non- Approved 6X in water; ssDNA 490 nm water at 4 oC ethidium bromide mutagenic in by MWRA 1 mL, $58 RNA Salmonella for drain TA98 and disposal, Dose : 5µL TA1537 April per 100 mL strains, w/ and 2010. agarose = w/o S9 $0.29 per activation. gel LD 50 , mouse subcutaneous : 250 mg/kg Aquatic toxicity : LC 50 not stated GelStar 6 dsDNA 493 nm agarose 20pg for Lasts 12 302nm or 312 Mutagenicity Treat as 10,000X in ssDNA 527 nm dsDNA; months nm filter for UV testing data potential DMSO, RNA acrylamide 25pg for when stored transilluminator not provided mutagen $158 for ssDNA; at -20 oC. per 2x250µL pre-cast 10pg for GelStar filter for LD 50 , rat( oral) : direction (agarose) RNA. photographs not stated of product Dose : DNA, maker. 4µL per 6 Cambrex QUESTIONS??? Send comments to [email protected] 5 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity post-gel Aquatic 40mL gel = toxicity : 125 LC 50 not stated gels@$1.27 per gel RNA, 8µL per 40mL gel = 62.5 gels@ $2.53 per gel. QUESTIONS??? Send comments to [email protected] 6 .
Load more

Recommended publications
  • Agarose Gel Electrophoresis
    Laboratory for Environmental Pathogen Research Department of Environmental Sciences University of Toledo Agarose gel electrophoresis Background information Agarose gel electrophoresis of DNA is used to determine the presence and distinguish the type of nucleic acids obtained after extraction and to analyze restriction digestion products. Desired DNA fragments can be physically isolated for various purposes such as sequencing, probe preparation, or for cloning fragments into other vectors. Both agarose and polyacrylamide gels are used for DNA analysis. Agarose gels are usually run to size larger fragments (greater than 200 bp) and polyacrylamide gels are run to size fragments less than 200 bp. Typically agarose gels are used for most purposes and polyacrylamide gels are used when small fragments, such as digests of 16S rRNA genes, are being distinguished. There are also specialty agaroses made by FMC (e.g., Metaphor) for separating small fragments. Regular agarose gels may range in concentration from 0.6 to 3.0%. Pouring gels at less or greater than these percentages presents handling problems (e.g., 0.4% agarose for genomic DNA partial digests requires a layer of supporting 0.8% gel). For normal samples make agarose gels at 0.7%. The chart below illustrates the optimal concentrations for fragment size separation. The values listed are approximate and can vary depending on the reference that is used. If you do not know your fragment sizes then the best approach is to start with a 0.7% gel and change subsequently if the desired separation is not achieved. Nucleic acids must be stained prior to visualization. Most laboratories use ethidium bromide but other stains (e.g., SYBR green, GelStar) are available.
    [Show full text]
  • Gelred® and Gelgreen® Safety Report
    Safety Report for GelRed® and GelGreen® A summary of mutagenicity and environmental safety test results from three independent laboratories for the nucleic acid gel stains GelRed® and GelGreen® www.biotium.com General Inquiries: [email protected] Technical Support: [email protected] Phone: 800-304-5357 Conclusion Overview GelRed® and GelGreen® are a new generation of nucleic acid gel stains. Ethidium bromide (EB) has been the stain of choice for nucleic acid gel They possess novel chemical features designed to minimize the chance for staining for decades. The dye is inexpensive, sufficiently sensitive and very the dyes to interact with nucleic acids in living cells. Test results confirm that stable. However, EB is also a known powerful mutagen. It poses a major the dyes do not penetrate latex gloves or cell membranes. health hazard to the user, and efforts in decontamination and waste disposal ultimately make the dye expensive to use. To overcome the toxicity problem In the AMES test, GelRed® and GelGreen® are noncytotoxic and of EB, scientists at Biotium developed GelRed® and GelGreen® nucleic acid nonmutagenic at concentrations well above the working concentrations gel stains as superior alternatives. Extensive tests demonstrate that both used in gel staining. The highest dye concentrations shown to be non-toxic dyes have significantly improved safety profiles over EB. and non-mutagenic in the Ames test for GelRed® and GelGreen® dyes are 18.5-times higher than the 1X working concentration used for gel casting, and 6-times higher than the 3X working concentration used for gel staining. This Dye Design Principle is in contrast to SYBR® Safe, which has been reported to show mutagenicity At the very beginning of GelRed® and GelGreen® development, we made a in several strains in the presence of S9 mix (1).
    [Show full text]
  • Supplementary Data for Publication
    Electronic Supplementary Material (ESI) for Physical Chemistry Chemical Physics. This journal is © the Owner Societies 2016 Supplementary Data for Publication Synthesis of Eucalyptus/Tea Tree Oil Absorbed Biphasic Calcium phosphate-PVDF Polymer Nanocomposite Films: A Surface Active Antimicrobial System for Biomedical Application Biswajoy Bagchi1,δ, Somtirtha Banerjee1, Arpan Kool1, Pradip Thakur1,2, Suman Bhandary3, Nur Amin Hoque1 , Sukhen Das1+* 1Physics Department, Jadavpur University, Kolkata-700032, India. 2Department of Physics, Netaji Nagar College for Women, Kolkata-700092, India. 3Division of Molecular Medicine, Bose Institute, Kolkata-700054, India. +Present Address: Department of Physics, Indian Institute of Engineering Science and Technology, Shibpur, Howrah, West Bengal-711103, India. §Present Address: Fuel Cell and Battery Division, Central Glass and Ceramic Research Institute, Kolkata-700032, India. *Corresponding author’s email id: [email protected] Contact: +919433091337 Antimicrobial activity of EU and TTO treated films on E .coli and S. aureus by acridine orange/ethidium bromide (AO/EB) dual staining Live/dead cell characterization of EU/TTO film treated bacterial cultures was also done to visualize the viability under fluorescence microscope (). The treated culture suspensions after 12 and 24 hours of incubation were collected by centrifugation (5000 rpm, 20 mins). The cell pellets were resuspended in PBS. The staining solution was prepared by mixing equal parts of acridine orange (5mg/mL) and ethidium bromide (3mg/mL) in ethanol. 20μL of the staining solution is then mixed with 10μL of the resuspended solution and incubated for 15 minutes at 37°C. 10μL of this solution was then placed on a glass slide and covered with cover slip to observe under fluorescence microscope.
    [Show full text]
  • SYBR Safe Case Study
    Replacing Ethidium Bromide in an Undergraduate Laboratory: SYBR Safe® Case Study March 2006 What is Ethidium Bromide and Why is it Used? Ethidium bromide (CAS #1239-45-8), or C21H20BrN3, is used in a number of laboratories, including those at MIT, for identifying DNA bands in samples that are loaded onto agarose gels. Ethidium bromide, commonly referred to as EtBr, binds to DNA. When placed under ultraviolet light, the EtBr-stained DNA bands fluoresce, allowing for the identification and visualization of nucleic acid bands. Ethidium bromide is considered an effective and relatively inexpensive technique for visualizing nucleic acid bands. Drawbacks of Ethidium Bromide Though effective and relatively inexpensive, ethidium bromide does have the following drawbacks for those handling the material in the lab: • it can be absorbed through the skin, irritating the eyes, mouth, and upper respiratory tract; • because of its tendency to intercalate in DNA bands, ethidium bromide is a powerful mutagen; • if handled indiscriminately in the lab, ethidium bromide can easily contaminate a large work area. When lab spaces are prepared for a move or for renovation, the space must be decontaminated of chemical, biological and radiological hazards. Because individual laboratories bear most, if not all, of the cost of decontaminating a lab, widespread ethidium bromide contamination may unnecessarily increase either the time or cost of lab preparation for moves or renovations; and • techniques for managing ethidium bromide waste are expensive - from a materials perspective, labor perspective, or both - or they beget more waste. Management of Ethidium Bromide Waste The United States Environmental Protection Agency (EPA) does not currently regulate ethidium bromide as a hazardous waste.
    [Show full text]
  • Ethidium Bromide Use and Disposal
    Ethidium Bromide Use and Disposal NOTE: Ethidium bromide is a chemical and should NOT be treated or labeled as a biohazard Ethidium Bromide (EtBr), commonly used in research laboratories as a stain for the visualization of nucleic acids in electrophoresis gels, is a toxic chemical and a potent mutagen. When used in nucleic acid staining, ethidium bromide fluoresces a red-orange to pink color under ultraviolet light and with increased fluorescence when bound to double-stranded DNA. While it is not specifically regulated as a hazardous waste, the mutagenic properties may present health hazards and disposal concerns if it is not managed properly in the laboratory. Required PPE: Always wear a lab coat, gloves, and appropriate protective eyewear when handling ethidium bromide and/or ethidium bromide containing material. Proper skin and eye protection are also needed when an ultraviolet (UV) light source is used while working with ethidium bromide. Avoid exposing unprotected skin and eyes to intense UV sources. A face shield is suggested if the UV source is pointing upwards. Guidelines for Ethidium Bromide Disposal: Stock Solutions: Stock solutions of ethidium bromide typically contain higher concentrations of ethidium bromide (approximately 10 mg/ml). Contact the Office of Environmental Safety & Services for collection and proper disposal of all unwanted stock solutions of ethidium bromide. Electrophoresis Gels and Buffers: Solid ethidium bromide waste (e.g., gels) typically contains 3 –5 ug/ml of ethidium bromide. Liquid ethidium bromide
    [Show full text]
  • Come-Back of Phenanthridine and Phenanthridinium Derivatives in the 21St Century
    Come-back of phenanthridine and phenanthridinium derivatives in the 21st century Lidija-Marija Tumir, Marijana Radić Stojković and Ivo Piantanida* Review Open Access Address: Beilstein J. Org. Chem. 2014, 10, 2930–2954. Laboratory for Study of Interactions of Biomacromolecules, Division of doi:10.3762/bjoc.10.312 Organic Chemistry and Biochemistry, Ruđer Bošković Institute, Bijenička cesta 54, PO Box 180, HR-10002 Zagreb, Croatia Received: 17 July 2014 Accepted: 21 November 2014 Email: Published: 10 December 2014 Ivo Piantanida* - [email protected] This article is part of the Thematic Series "Nucleic acid chemistry". * Corresponding author Guest Editor: H.-A. Wagenknecht Keywords: ds-DNA and ds-RNA binding; intercalation; minor groove binding; © 2014 Tumir et al; licensee Beilstein-Institut. nucleic acids; organic synthesis; phenanthridine; phenanthridinium License and terms: see end of document. Abstract Phenanthridine derivatives are one of the most intensively studied families of biologically active compounds with efficient DNA binding capability. Attracting attention since DNA structure discovery (1960s), they were early recognized as a symbol of DNA intercalative binding, for many decades applied as gold-standard DNA- and RNA-fluorescent markers (ethidium bromide), probes for cell viability (propidium iodide), but also “ill-famed” for various toxic (genotoxic) and mutagenic effects. After two decades of low interest, the discovery of phenanthridine alkaloids and new studies of antiparasitic/antitumor properties of phenanthridine derivatives resulted in the strong increase of the scientific interest about the turn of this century. Here are summarized phenanthri- dine-related advances in the 21st century (2000-present period) with emphasis on the supramolecular interactions and bioorganic chemistry, as well as novel or improved synthetic approaches.
    [Show full text]
  • Ethidium Bromide: Disposal, Decontamination, Procedure: 8.03 Created: 2/28/2012 and Destruction Version: 2.1 Revised: 1/27/2016
    Ethidium Bromide: Disposal, Decontamination, Procedure: 8.03 Created: 2/28/2012 and Destruction Version: 2.1 Revised: 1/27/2016 A. Purpose 1. This policy provides general guidance to researchers and labs on how to work safely with Ethidium bromide. This mix has been designated as “particularly hazardous” by OSHA. This Policy will describe the minimum requirements for the safe storage, use, handling, and disposal of particularly hazardous substances, including spill and accident response procedures. 2. Ethidium bromide is mutagenic and moderately toxic and must be handled with care. The powder form is considered an irritant to the upper respiratory tract, eyes, and skin. Ethidium bromide is thought to act as a mutagen because it bonds in double stranded DNA, thereby deforming the molecule. This is believed to block or trip biological processes occurring on DNA. Preparation of stock solutions and any operations capable of generating Ethidium bromide dust or aerosols should be conducted in a fume hood to prevent inhalation. Nitrile gloves, a lab coat, and eye protection should be worn at all times, as with working with any hazardous material. B. Applicability/Scope 1. This Policy is applicable to, and must be adhered to by, all Columbia University laboratory workers (i.e., Principal Investigators, laboratory personnel, students, visiting researchers, etc.) who use or work with Ethidium bromide. Careful handling and stringent controls of Ethidium bromide is essential in order to protect workers and the environment, and to comply with OSHA regulations. 2. Additional safety requirements may apply, depending on if Ethidium bromide is mixed/used with a specific chemical.
    [Show full text]
  • SYBR® Green Staining Reagent, DNA Free
    SYBR® Green staining reagent, DNA free 10x concentrated SYBR® Green I staining solution, DNA-free Product No. A8511 Description SYBR® Green is an asymmetrical cyanine dye. It is used as intercalating dye for the general detection of double-stranded DNA (dsDNA). Our 10-fold concentrated DNA-free SYBR® Green I dye solution is particularly suitable for qPCR using general primers such as 16S rDNA or 18S rDNA primers. An additional application is the staining of DNA in gel electrophoresis. SYBR® Green shows lower mutagenic potential in comparison to ethidium bromide [1]. Thus, SYBR® Green is often used as a substitute to the classical Ethidium bromide dye. Nevertheless, follow the usual safety precautions dealing with DNA dyes. Synergistic effects have been shown to increase mutagenicity of the dye [2]. The complex of DNA and SYBR® Green absorbs blue light of wavelength 494 nm (absorption maximum) and emits green light at 521 nm (emission maximum). The stained DNA can be detected on a blue light transilluminator. Other absorption maxima in the UV range are at 284 nm and 382 nm. Hence, SYBR Green- stained DNA can also be detected on the UV transilluminator. Available pack sizes: Article No. A8511,10625 1 vial of 0.625 ml Article No. A8511,50625 5 vials of 0.625 ml Article No. A8511,100625 10 vials of 0.625 ml Literature: [1] Singer VL, Lawlor TE, Yue S. (1999) Comparison of SYBR Green I nucleic acid gel stain mutagenicity and ethidium bromide mutagenicity in the Salmonella/mammalian microsome reverse mutation assay (Ames test). Mutation Research 439: 37-47.
    [Show full text]
  • PGMD/Curcumin Nanoparticles for the Treatment of Breast Cancer Mankamna Kumari1, Nikita Sharma1, Romila Manchanda2, Nidhi Gupta3, Asad Syed4, Ali H
    www.nature.com/scientificreports OPEN PGMD/curcumin nanoparticles for the treatment of breast cancer Mankamna Kumari1, Nikita Sharma1, Romila Manchanda2, Nidhi Gupta3, Asad Syed4, Ali H. Bahkali4 & Surendra Nimesh1* The present study aims at developing PGMD (poly-glycerol-malic acid-dodecanedioic acid)/curcumin nanoparticles based formulation for anticancer activity against breast cancer cells. The nanoparticles were prepared using both the variants of PGMD polymer (PGMD 7:3 and PGMD 6:4) with curcumin (i.e. CUR NP 7:3 and CUR NP 6:4). The size of CUR NP 7:3 and CUR NP 6:4 were found to be ~ 110 and 218 nm with a polydispersity index of 0.174 and 0.36, respectively. Further, the zeta potential of the particles was − 18.9 and − 17.5 mV for CUR NP 7:3 and CUR NP 6:4, respectively. The entrapment efciency of both the nanoparticles was in the range of 75–81%. In vitro anticancer activity and the scratch assay were conducted on breast cancer cell lines, MCF-7 and MDA-MB-231. The IC50 of the nanoformulations was observed to be 40.2 and 33.6 μM at 48 h for CUR NP 7:3 and CUR NP 6:4, respectively, in MCF-7 cell line; for MDA-MB-231 it was 43.4 and 30.5 μM. Acridine orange/EtBr and DAPI staining assays showed apoptotic features and nuclear anomalies in the treated cells. This was further confrmed by western blot analysis that showed overexpression of caspase 9 indicating curcumin role in apoptosis. With the changing environmental factors and lifestyle such as pollution, tobacco smoking, diet patterns, there has been a tremendous increase in the incidence of cancer.
    [Show full text]
  • Ethidium Bromide(CAS#1239-45-8) ) Frequently EH&S Facts
    EH&S Facts Fact Sheet Fact Ethidium Bromide (CAS# 1239-45-8) What is Ethidium Bromide? Ethidium bromide (EtBr) is commonly used as a non-radioactive DNA stain to identify and visualize nucleic acid bands in electrophoresis and perform other methods of nucleic acid separation. Solutions of EtBr fluoresce readily with a reddish-brown color when exposed to ultraviolet (UV) light. Although it is an effective tool for genomic research, its hazardous properties require special safe handling and disposal. What are the potential health hazards of Ethidium Bromide? • EtBr is a mutagen (may cause genetic damage) and is moderately toxic after an acute exposure • EtBr can be absorbed through skin, and will stain it purple • EtBr is an irritant to the skin, eyes, mouth, and upper respiratory tract • Some alternative stains are less mutagenic and less toxic than EtBr. If the toxicological data is lacking or unclear, handle the stain in the same way as EtBr • Some alternative stains are suspended in dimethyl sulfoxide (DMSO), which can increase skin absorption of organic compounds What special laboratory precautions are recommended? Good laboratory work practices help reduce hazardous exposures. 3 Use dry materials in a fume hood, or 3 Review Material Safety Data Sheet choose premixed solutions to avoid (MSDS) and this EH&S fact sheet before inhalation exposure handling DNA stains 3 Wear nitrile gloves, a laboratory coat, 3 Ensure unobstructed access to an safety glasses, long pants, and closed- emergency eyewash/shower unit in the toed shoes. Change
    [Show full text]
  • Interaction Between DNA, Albumin and Apo-Transferrin and Iridium(III) Complexes with Phosphines Derived from Fluoroquinolones As a Potent Anticancer Drug
    pharmaceuticals Article Interaction between DNA, Albumin and Apo-Transferrin and Iridium(III) Complexes with Phosphines Derived from Fluoroquinolones as a Potent Anticancer Drug Sandra Amanda Kozieł 1,* , Monika Katarzyna Lesiów 1, Daria Wojtala 1, Edyta Dyguda-Kazimierowicz 2 , Dariusz Bie ´nko 2 and Urszula Katarzyna Komarnicka 1,* 1 Faculty of Chemistry, University of Wroclaw, Joliot-Curie 14, 50-383 Wroclaw, Poland; [email protected] (M.K.L.); [email protected] (D.W.) 2 Faculty of Chemistry, Wroclaw University of Science and Technology, Wybrzeze˙ Wyspia´nskiego27, 50-370 Wroclaw, Poland; [email protected] (E.D.-K.); [email protected] (D.B.) * Correspondence: [email protected] (S.A.K.); [email protected] (U.K.K.) Abstract: A group of cytotoxic half-sandwich iridium(III) complexes with aminomethyl(diphenyl) phosphine derived from fluoroquinolone antibiotics exhibit the ability to (i) accumulate in the nucleus, (ii) induce apoptosis, (iii) activate caspase-3/7 activity, (iv) induce the changes in cell cycle leading to G2/M phase arrest, and (v) radicals generation. Herein, to elucidate the cytotoxic effects, we investigated the interaction of these complexes with DNA and serum proteins by gel electrophoresis, fluorescence spectroscopy, circular dichroism, and molecular docking studies. DNA binding experiments established that the complexes interact with DNA by moderate intercalation Citation: Kozieł, S.A.; Lesiów, M.K.; and predominance of minor groove binding without the capability to cause a double-strand cleavage. Wojtala, D.; Dyguda-Kazimierowicz, The molecular docking study confirmed two binding modes: minor groove binding and threading E.; Bie´nko,D.; Komarnicka, U.K.
    [Show full text]
  • Redsafe Technical Bulletin Eng V1.0
    RedSafe Technical Bulletin │ Jul, 2017 (1st Edition) ISO 9001 / 14001 / 13485 Certified Company Technical Bulletin Characteristics and Usage of RedSafe Nucleic Acid Staining Solution (20,000X) Abstract RedSafe Nucleic Acid Staining Solution (20,00x) is a new and safe alternative favourably reviewed by many researchers against ethidium bromide(EtBr) which is a generally accepted traditional method when dyeing nucleic acid in Agarose gel. This manual aims to secure the best result by guiding right usage and the characteristics of RedSafe as materials, to be a guideline of efficient use of the product by providing answers for many common questions. RedSafe has its excitation not only at ultraviolet wave such as UV-A (especially 270nm) and UV-B (300nm), but also at the visible light wave range of 420 ~ 510 nm. RedSafe can be compatible with UV Transilluminator that are generally diffused and also compatible with Blue LED (generally 420nm) known as an alternative for UV recently. RedSafe that drained energy from each wave emits strong green florescence at 520 ~ 550nm (exactly at 535nm) so that it can help DNA analysis after electrophoresis. Electrophoresis DNA staining Ethidium bromide is an intercalating agent commonly used as a Electrophoresis is the motion of dispersed particles relative to a fluid fluorescent tag (nucleic acid stain) in molecular biology laboratories under the influence of a spatially uniform electric field. This electro for techniques such as agarose gel electrophoresis. It is commonly kinetic phenomenon was observed for the first time in 1807 by abbreviated as "EtBr", which is also an abbreviation for bromoethane. Ferdinand Frederic Reuss (Moscow State University), who noticed When exposed to ultraviolet light, it will fluoresce with an orange that the application of a constant electric field caused clay particles colour, intensifying almost 20-fold after binding to DNA.
    [Show full text]