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Identification of Proteins in Urine Samples by Cellulose Acetate Electrophoresis (CAT#: STEM-ET-0310-ZJF)

Introduction

High‐throughput mass spectrometric analysis of urinary proteins is being widely used. Since urine sampling is noninvasive and relatively easy compared with serum samples, urine is attractive for biomarker discovery. A urinary protein panel, which is a renal disease assessment involving several novel protein markers, increases the diagnosis accuracy in diabetes mellitus, lupus nephritis, and recurrent bladder cancer. Therefore, a multiplex urinary protein assay is valuable for monitoring renal diseases. Cellulose acetate electrophoresis is one of the most classic approaches of protein panel analysis.<br />In this service, proteins would be extracted from each band fractionated by CAE and analyzed by mass spectrometry. Analysis of urinary proteins by CAE is a useful and challenging method for the recognition of damaged sites in the kidney.




Principle

Cellulose Acetate Electrophoresis is one of the zone electrophoresis and the supporting medium of it is cellulose acetate menbrance. Cellulose acetate is derived from the acetylation of the cellulose hydroxyl group. Under an electric field, the sample migrates on the cellulose acetate film in the buffer system based on the charge.
Although its resolution is lower than that of polypropionamide gel electrophoresis (PAGE), its features of simplicity, rapidity and adaptability make it retain its place in routine clinical diagnosis.

Applications

Clinical analysis, proteome, urinary protein, kidney

Procedure

1. Preparation: Prepare electrophoresis buffer with a certain pH. Soak suitable-sized cellulose acetate films in the buffer solution for some time. Then use filter paper to absorb the excess buffer on the surface.
2. Sample Application: Apply a certain amount of sample in a strip on the cellulose film with slides or capillary tubes.
3. Electrophoresis: Add an appropriate amount of buffer to the electrophoresis tank to soak the platinum electrode. Cut wicks from filter paper or chromatography paper and place them along the bridges in the tank. Place cellulose films on filter paper wicks, the sample strips near the cathode. Switch on the electrophoresis apparatus and set the voltage and current. Perform electrophoresis for a period of time.
4. Determination: Stain the film and bleach it until the background is colorless. Blow dry the film and cut the separated strips. Put the strips in the liquid elution in test tubes to treat them. Finally, measure the absorbance of each tube.

Materials

• Cellulose acetate electrophoresis apparatus
• Cellulose acetate menbrance
• Sample solution
• Buffer solution