HOME       

Cellogel/Myl is a wet Cellogel membrane supported on Mylar. Cellogel/Myl was produced to satisfy the requests from laboratories using the Helena system of “horizontal electrophoresis” on dry acetate. This can, in fact, substitute Titan III in Zip Zone chambers with the advantage of practicality and the possibility to perform high resolution electrophoresis. Cellogel/Myl facilitates the discovery of monoclonal bands because it permits the lengthening of the gamma zone much more than agarose and other known gels, thanks to the strong electroendosmosis with retromigration of the immunoglobulins. It is, therefore, suitable not only for high resolution electrophoresis but also for high definition electrophoresis as shown in the example illustrated where the gamma zone extends for about 7 cm with the polyclonal immunoglobulins reduced to a minimum and the small monoclonals compacted and enhanced to the maximum.

High Definition Electrophoresis on Cellogel/Myl is an advanced system of High Resolution for serum protein, urine, cerebrospinal fluid, etc. proposed by Cellogel Co. for the clinical identification of difficult cases of monoclonal gammapathies and also for the identification of the protein “Minor Components” of serum. With this method 20-30 bands can be revealed with only 0.9 µl of specimen. Two procedures are proposed for immunovisualization of the bands:

Rollblotting with immediate transfer of all the separated components onto a membrane of nitrocellulose by the method of Cohen, Lambrey and Dropsy (J. Imm. Meth. 104, 25-30, 1987). The nitrocellulose membrane is subsequently incubated with mixtures of specific monoclonal antibodies in a 1% BSA solution (blocking). With this procedure the Minor Components present in quantities of a few picograms can be identified. Immunovisualization is performed with the system of Streptavidin biotin HRP (code 926 MA) suitable for revealing antibodies, both monoclonal and polyclonal of rabbit. It is possible to reveal tumour markers as, for example, CEA, Ferritin, AFP, PSA, beta2-micro etc.

Buffer the film in Tris Hippurate (code 02C13-2X-6) for 10 minutes in minibox (code 13A51) on rotating shaker; 20 ml of buffer solution are sufficient. Horizontal buffering gives rise to savings of large volumes of buffer. The same can be said of staining and destaining. During buffering pour the buffer solution into the tank (200 ml per compartment), wet the paper wicks (code 13B19) in the tank and place it astride the bridge.
Cellogel/Myl has the advantage that it can be immersed directly into the buffer as it is already wet and does not present the problem of entrapping air in its pores as occurs with dry acetate which is normally buffered vertically.

Pipette 20 µl of serum on each drop holder of the specimen holder base. Check that the applicator is perfectly clean.

Remove excess buffer solution between 2 sheets of filter paper and place the film on the base of the applicator. Apply the specimens at about 3.5 cm from the cathodic edge for 10 seconds.

Position the film in the tank with the gel facing downwards and the deposit zone at the negative pole. Ascertain that contact between the film and the paper wicks of the bridge is perfect.

Plug the tank into the power supply, cover the tank and apply 160 V for 18 minutes to obtain 22 mm migration.

After electrophoresis, switch off the power supply. Extract the strip and immerse it in a bath of Ponceau S for 5 minutes. Return the stain to its bottle. Destain in acetic acid 5% (or 3% odourless citric acid) for 3 baths of 3 - 4 minutes each. Clearing: immerse in a bath of clearing solution (code 06A06-S1) for 5 minutes. Remove excess liquid with a glass rod and heat in thermostat at 80°C for 10 minutes. After cooling for 1 minute at room temperature the film is placed on the scanner for the densitometric reading.

With Cellogel/Myl it is possible to perform electrophoresis for:
- Research
- Immunological techniques, such as Ouchterlony Technique and the Mancini Technique of Quantitative Radial Immunodiffusion.