PurMa Ethidum Bromide Solution 10 mg/ml - PurMabiologics

PurMa Ethidum Bromide Solution 10 mg/ml

$5.52$157.42

Ethidium bromide is a sensitive fluorescent dye that commonly used as a staining agent for nucleic acid detection through gel electrophoresis techniques in molecular biology laboratories. It can detect both single and double-stranded nucleic acids (both DNA and RNA), However, Ethidium Bromide does not intercalate into single-stranded DNA very well, thus fluorescence of single-stranded DNA is relatively poor in comparison to double-stranded DNA. That’s why double-stranded DNA from PCRsrestriction digests, etc. are usually detected through ethidium bromide. It’s strong ability to bind itself with DNA by intercalating between DNA base pairs of the double helix, makes ethidium bromide a powerful inhibitor of DNA polymerase. It is also used with acridine orange (fluorescence dye used to stain acidic vacuoles, DNA and RNA in living cells) to differentiate between viable, apoptotic and necrotic cells.

Description

Ethidium bromide is a sensitive fluorescent dye that commonly used as a staining agent for nucleic acid detection through gel electrophoresis techniques in molecular biology laboratories. It can detect both single and double-stranded nucleic acids (both DNA and RNA), However, Ethidium Bromide does not intercalate into single-stranded DNA very well, thus fluorescence of single-stranded DNA is relatively poor in comparison to double-stranded DNA. That’s why double-stranded DNA from PCRsrestriction digests, etc. are usually detected through ethidium bromide. It’s strong ability to bind itself with DNA by intercalating between DNA base pairs of the double helix, makes ethidium bromide a powerful inhibitor of DNA polymerase. It is also used with acridine orange (fluorescence dye used to stain acidic vacuoles, DNA and RNA in living cells) to differentiate between viable, apoptotic and necrotic cells.

Ethidium bromide aqueous solution has UV absorbance maxima at 210 nm and 285 nm. As a result of this excitation, Ethidium bromide solution emits orange light with 605 nm wavelength which called emission maximum.

PurMabiologics LLC provides ready- to- use Ethidium Bromide solution, Molecular Grade for most of the molecular biology applications of fluorometric nucleic acid assays (Specially PAGE and agarose gel electrophoresis. Safe and convenient Ethidium bromide solution is supplied as 10mg/ml aqueous solution in various convenient sizes. The solution can be used with dilution to your desired concentration or as provided. The economic standard 10 mg/ml stock solution is prepared in deionized water that provides advantage in reducing hazard exposure.

Application

  • Staining for Polyacrylamide gel electrophoresis (PAGE)
  • Staining for Agarose gel electrophoresis (AGE)
  • To detect protein– DNA complexes in band shift assays
  • Large scale plasmid preparation using cesium chloride gradients
  • To detect polymerase chain reaction products
  • As a stain to visualize cells to assess cell viability

 

Specifications:

Staining agent  Ethidium bromide
Cas Number 1239-45-8
Molecular Formula C21H20BrN3
Chemical structure
Molecular Weight 394.31
synonym 5-ethyl-6-phenylphenanthridin-5-ium-3,8-diamine bromide
Color Purple-Red
Physical state Liquid
Concentration 10mg/ml in water
Purity: >98.0%
Solubility Soluble in water, and 750 parts chloroform at 20 °C.

 

Boiling Point 100° C

 

Fluorescence λex 480 nm, λem 620 nm in water

 

pK Values pKb: 1.67 (Predicted)

 

Target Molecule DNA, RNA
Detection Method Fluorescent
Detection Location In-Gel Detection
Sensitivity Can detect as little as 1 ng nucleic acid in agarose gels
DNase/RNase Assay None detected
Phosphatase Assay None detected
Protease Assay None detected
Shipping condition Room Temperature

Product Highlights:

  • Molecular biology grade
  • Highly sensitive- Can detect as little as 0.5 ng of nucleic acids in agarose gels
  • Versatile – Can detect both single and double-stranded DNA or RNA, can be used with acridine orange.
  • Convenient -Can be used as-it-is or diluted to the desired concentration
  • Safe -Aqueous form reduces the chance for hazard exposure of ethidium bromide

Protocol for Gel Staining

After electrophoresis:

  1. To make 200 ml working solution, simply add 10 µL of 10 mg/mL ethidium bromide solution to 200 mL ddH2O. To make 1000 ml working solution, simply add 50µl of 10mg.ml Ethidium bromide solution. The dilution makes ideal concentration of 0.5 µg/mL gel staining working solution
  2. Soak the gel with working solution as needed to merge it for 15-30 minutes with gentle agitation. Longer staining times will result in high background.
  3. Rinse the gel with ddH2O
  4. De-stain the Gel with fresh ddH2O for 15 minutes with gentle agitation to decrease background.
  5. If necessary, another 15 mins the gel can be de-stained for better picture
  6. After de-staining, the gel will be ready for photographing by DNA gel imager, or for visualization of DNA bands by UV light box (254 nm wavelength)

Before Electrophoresis:

Ethidium bromide can also be incorporated with melted agarose gel or in running buffer at 0.5μg/ml before running the electrophoresis and visualized immediately after electrophoresis. The detection limit of DNA bound to ethidium bromide is 0.5 to 5.0 ng/band.

Caution:

Ethidium bromide is known as a frameshift mutagen. It is also a toxic chemical causes eye and skin irritation. Always wear gloves while dealing with this chemical if the skin is contaminated, wash with plenty of water. Follow the MSDS for proper handling.

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