Gel Electrophoresis

Cellulose Acetate Electrophoresis

High Resolution Ampholytes for Isoelectric Focussing

Cellulose Acetate Electrophoresis

Cellulose acetate electrophoresis nowadays plays an important role in clinical diagnostics routine procedures, but has also helped to investigate a broad range of subjects in life science research (see applications).

Cellulose acetate electrophoresis seperates proteins primarily by charge. Protein migration takes place on the buffer film on the surface of the cellulose acetate plate. The buffer used depends on the enzyme being screened. Cellulose acetate plates require less sample and stain volume and allow shorter gel running and staining time than other mediums.

Applications

Dry Cellulose Acetate Strips

Humid Cellulose Acetate Strips

List of Cellulose Acetate Strips (dry and humid)

Non-toxic Reagents Systems

Applications

Analysis of hemoglobin is a typical example, there cellulose acetate electrophoresis is superior to other methods. Other examples are the separation of enzymes (e.g. creatinine phosphokinase, GOT, acidic erythrocyte phosphatase, phosphoglucomutase, etc.), muco-polysaccharides, plasma, serum, cerebrospinal fluid, urine and other body fluids. Another field of applications is the quality control of biological compounds. Cellulose acetate electrophoresis has been shown to be an accurate, but simple, method for protein quantification.

References

(1) Goldbloom R.B., Screening for hemoglobinopathies in Canada, in Canadian Task Force on the Periodic Health Examination. Canadian Guide to Clinical Preventive Health Care. Ottawa: Health Canada (1994), p. 206 - 218.

(2) Barufaldi, M.; Pappano, N. B.; Debattista, Nora B., Quantitative Protein Determination from Cellulose Acetate Strip Electrophoresis, J. Chem. Educ., 76 (1999), p. 965

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Dry Cellulose Acetate Strips

With our dry cellulose acetate strips you can separate nearly all sample materials that migrate in an electrical field. You can use substance mixtures of 0.06 to 0.15 mg per cm application line. The start concentration of the mixture to be separated should be more than 1 mg/ml. Weaker solutions should be enriched or concentrated. Advantages of those strips include:

• High porosity of the strip (approx. 80 % free pore volume)
• Isotropic, integral matrix for ideal flow conditions
• Minimal diffusion effect and low adsorption capacity
• Perfect transparency because of the pore system
• Ideal for the non-toxic ATX reagent system
• Virtually unlimited stability at room temperature
• Two identical sides, no need to search for the "right" side
• Available in different sizes for all current chamber types

Besides standard cellulose acetate electrophoresis those strips can also be used for high-voltage electrophoresis, chromatography and immunodiffusion.

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Humid Cellulose Acetate Strips

The humid state ensures that strips are rapidly buffered and that gel properties are maintained. Owing to a unique structure (conical pores) these strips are an ideal supporting medium for both analytical and micro-analytical electrophoresis and for all immunological techniques. Thus they are a genuine alternative to agarose gels providing results at least as good as with agarose gels. Compared with dry cellulose acetate strips, the wet ones have the following advantages:

• The samples do not run into each other even during multiple application.
• These strips reliably absorb the samples and guarantees optimal application, even of a large sample volume
• Even with low-voltage electrophoresis, wet strips guarantee sharp bands. There is no over-lapping.
• Owing to their special structure, there is no streaking. Endosmotic effects are safely avoided, especially for important immunological techniques. Thus you will receive better and more reproducible results in immunological techniques.
• Lower costs, less time efforts.
• They are extremely resistant to strong alkalis, which means, for example, that staining with conventional oil-soluble sudan dye in NaOH causes no problem.

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Table 1: Cellulose Acetate Strips, Product List

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Non-toxic Reagents Systems

With conventional electrophoresis reagents, there is always a risk to health. By contrast, the reagents in the ATX reagent system are non-toxic, environmentally friendly. Conventional reagents contain various toxic substances: buffer solutions contain barbituric acid, classified as a narcotic according to the Dangerous Drugs Act; staining solutions contain methanol and acetic acid which irritate the skin and airways; clarifying solutions contain methanol and dioxan isobutanol, a carcinogenic, inflammable substance which is classified as hazardous waste.

Stop using these toxic substances. For your electrophoresis, start using the non-toxic ATX reagent system and protect yourself and the environment against permanent damage.

Advantages include:

• ATX-reagents are non-toxic and are non-irritant to skin and airways
• ATX-reagents are environmentally friendly
• ATX-reagents pose no disposal problems
• ATX-reagents are odourless
• ATX-reagents are non-inflammable
• ATX-reagents are stable at room temperature
• ATX-reagents are specially coordinated with ATX micro-solid strips
• ATX-reagents are available as ready-to-use solutions or as concentrates
• ATX-reagents guarantee excellent, reproducible results

Table 2: ATX Reagents, Product List

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High Resolution Ampholytes for Isoelectric Focussing

Isoelectric focusing is an electrophoretic separation based on isoelectric point of proteins. It is used as alternative electrophoresis format completing the widely used SDS-PAGE electrophoresis, which is based on size of proteins. Specific separation problems, such as the differentiation of protein isoforms or enantionmeres, have been successfully solved by isoelectric focusing (IEF).^[1] That makes it a very useful technique especially for purposes like food analysis, species determination, or seed testing.

Compared to alternative methods (e.g. PCR based), IEF is:

• efficient
• expressive
• economic
• easy
• fast

[1] P. Glukhovskiy, G. Vigh, Analytical and Preperative Scale Isoelectric Focusing Separation of Enantiomers, Anal. Chem., 71 (17), 1999, 3814 - 3820.

Carrier Ampholytes

Isoelectric focusing as stand alone analytical or micropreparative technique requires ampholytes, complex mixtures of bifunctional amphoteric (both acid and basic) buffer molecules which form a pH gradient in the medium during electrophoresis. Within that gradient proteins will migrate toward the anode or cathode until it arrives at the point equal to its pI value. At this point it will have a net charge of zero and is said to be focused. Carrier ampholytes have to be added to the separation medium (polyacrylamide, agarose, or sephadex). They are supplied as 40% solution and typically diluted to a final concentration of 2% in the IEF matrix.

Advantages of High Resolution Ampholytes

A new series of carrier ampholytes (see table 1) is based on a specific method to produce mixtures of numerous polyamino-polycarboxylic acids. These new carrier ampholytes have been developed for use of isoelectric focusing in quality control applications, especially in food and seed analysis. Their superior properties include:

• High buffer capacity: Compared to established ampholytes they offer buffering capacities 1.5-2.5 times higher (up to pH 8, equal above pH 8)
• Good even conductivity
• Linear and reproducible pH gradient
• High resolution

Table 1.

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