Electrophoresis Preparation and Sample Application on Acetate Cellulose Membrane

Preprocessing Acetate Cellulose Membrane

Cutting Membrane:

Cut the acetate cellulose membrane into specific sizes based on the quantity of separated samples, typically 2.5cmx11cm or 7.8cmx15cm.


Marking Sample Application Line:

  • On the non-glossy side of the membrane, lightly mark the sample application line with a pencil.
  • The location of the application line can be chosen 2-3cm from one end of the membrane or sometimes near the centerline.
  • The position of the application line is usually determined based on preliminary electrophoresis experiments.


Immersing in Electrode Buffer Solution:

  • In a shallow dish or culture dish, pour in electrode buffer solution.
  • Carefully float the membrane on the surface of the electrode buffer solution, ensuring the non-glossy side faces down.
  • As the bottom of the membrane absorbs the electrode buffer solution, it gradually sinks until fully submerged.

Removing and Absorbing Excess Liquid:

  • After the membrane has permeated the electrode buffer solution, use blunt forceps to carefully remove it.
  • Sandwich the membrane between two layers of filter paper to absorb excess electrode buffer solution, avoiding excessive dryness.
  • If a white opaque area appears on the membrane, indicating excessive drying, reimmerse the membrane in electrode buffer solution and absorb to the appropriate degree with filter paper.

Sample Application Process

Choosing and Operating on Sample Application Line:

Apply the sample along the sample application line on the non-glossy side of the membrane, usually in a linear pattern rather than a point-like application.

Qualitative Analysis Sample Application:

  • Use capillary tubes (with a diameter of 0.5~1.0mm) or molds for qualitative analysis sample application.
  • During qualitative analysis, dip the sample and “stamp” it along the sample application line.

Quantitative Analysis Sample Application:

  • Use a microliter syringe for quantitative analysis sample application.
  • When using a capillary tube or microliter syringe, smoothly move it up and down or left and right along the sample application line until the predetermined volume of the sample is applied.

Controlling Sample Line Dimensions:

  • After applying each sample on the membrane, the length of the sample line is generally 1.5cm, with a width generally not exceeding 4mm.
  • The distance between sample lines and between the sample line and the long edge of the membrane is generally 3~5mm.

Adjusting Sample Volume:

The amount or volume of the sample applied varies significantly depending on factors such as sample concentration, staining, and analytical methods.

Sample Volume during Quantitative Analysis:

For quantitative analysis, the applied sample volume on each centimeter of the sample application line is generally 0.1-0.5μl, equivalent to the amount of a 5-1000μg protein sample.


Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) has specialized in manufacturing electrophoresis instruments for more than 50 years with our own professional technical team and R&D center. We have reliable and complete production line from design to inspection, and warehouse, as well as marketing support. Our main products are Electrophoresis Cell (tank/chamber), Electrophoresis Power Supply, Blue LED Transilluminator, UV Transilluminator, Gel Image & Analysis System etc.

Liuyi Bilotechnology manufactures one model of electrophoresis tank called cellulose acetate membrane elctrophoresis tank DYCP-38C which can be adapted for the analysis of hemoglobin variants, including those associated with sickle cell disease (SCD). It is a traditional and economic method for hemoglobin electrophoresis.  


If you are interested in this simple and economic electrophoresis method, please visit the following article for protocols and guidelines of hemoglobin electrophoresis offered by our experimenters.

Experiment for separating serum protein by Cellulose Acetate Membrane Electrophoresis  

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Post time: Jan-30-2024