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 ¤  Materials and Me...
 ¤  Results
 ¤  Discussion
 ¤  Summary
 ¤  Acknowledgement
 ¤  References

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ORIGINAL CONTRIBUTION
Year : 1997  |  Volume : 51  |  Issue : 4  |  Page : 113-114
 

Reuse of cellulose acetate membrane strips for protein and haemoglobin electrophoretic analysis


Department of Clinical Biochemistry, Wanless Hospital, Miraj Medical Centre, Miraj-416 410, Maharashtra, India

Correspondence Address:
M M Satwekar
Department of Clinical Biochemistry, Wanless Hospital, Miraj Medical Centre, Miraj-416 410, Maharashtra
India
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PMID: 9355695

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How to cite this article:
Satwekar M M. Reuse of cellulose acetate membrane strips for protein and haemoglobin electrophoretic analysis. Indian J Med Sci 1997;51:113-4

How to cite this URL:
Satwekar M M. Reuse of cellulose acetate membrane strips for protein and haemoglobin electrophoretic analysis. Indian J Med Sci [serial online] 1997 [cited 2014 Nov 1];51:113-4. Available from: http://www.indianjmedsci.org/text.asp?1997/51/4/113/11546


Electrophoretic studies have unique and widely accepted role in the clinical chemistry laboratory. These investigations are usually referred to as "Special Investiga­tions" due to variety of reasons such as a) personnel training, b) cost of the operation and c) inter­pretation of the results obtained. Personnel training and interpreta­tion of results could be overcome by proper training but the cost of the Electrophoretic analysis be­comes the major limiting factor. [1] Cellulose acetate membrane (CAM) as a support medium for electrophoresis has been exten­sively used in almost all clinical laboratories This has major advan­tages over the other support media with regard to speed of separa­tion, simplicity of operation and good, prominent resolution of pro­tein zones . [2],[3],[4]

Over a period of last nine months we have analysed 60 specimens for protein and hemo­globin electrophoretic studies. Seven cases of Serum protein electrophoresis (SPE) SPE were confirmed as having monoclonal gamopathy while one case of sic­kel cell haemoglobinopathy was detected.


 ¤ Materials and Methods Top


The CAM were made available from Gelman sciences Inc, Ann Arbor, MI 48601. All other rea­gents used for this study were of analytical grade. Electrophoresis was run according to the standard procedure . [3] Once the electropho­retic analysis of either protein or haemoglobin was over the CAM were stained with 0.2% w/v Ponceau's prepared in 3% w/v richloroacetic acid. The staining was allowed for 30 to 45 minutes at room temperature followed by destaining with 5% v/v glacial ace­tic acid. If this CAM strip after staining and destaining revealed normal protein pattern on visual inspection, the same CAM strip was effectively reused for another electrophoretic analysis. The CAM strip after the electrophoretic run, staining and destaining was soak­ed in petridish containing 0.001 N NaOH for 20 to 30 minutes at room temperature, with intermittent shaking. This was further washed with double distilled water for 5 to 6 times, with an interval of 5 minutes. These washed CAM strips were dried on blotting paper and were soaked in the appropriate buffer used for electrophoresis, till further used.


 ¤ Results Top


The method used for cleansing the CAM strip with 0.001N NaOH is quite convenient, cheap and reliable approach to clear off all the protein zones from CAM strip. The strength of the alkali (NaOH) is kept low so that the microtexure and porosity of CAM strip remains intact and gives better results dur­ing further use.


 ¤ Discussion Top


The reuse of laboratory mate­rials is of interest both for econo­mic and ecological grounds. The high cost of the electrophoretic analysis many a times goes beyond the paying capacity of needy and poor patients. The successful re­use of CAM strip for subsequent electrophoretic analysis would help to reduce the cost. We have exa­mined electrophoretic pattern of seven cases of monoclonal gammo­pathv and we were successful in getting similar kind of electrojho­retic pattern by reusing the CAM strip for 6 to 7 times. Further re­use of CAM strip for subsequent analysis depends upon the quality of the individual strip, however, it can be reused effectively for at least 6 to 7 times.


 ¤ Summary Top


The exorbidant cost of electro­phoretic analysis, many a times be­comes the major limitation factor. As most of the clinical labora o­res import the cellulose acetate membrane which costs 5 to 6 U.S. that is Rs. 180/- to Rs. 210/-, the final cost goes up to Rs. 350/- to Rs. 500/-. We have observed that CAN strips can be effectively re­used 6 to 7 times for subsequent electrophoretic analysis.


 ¤ Acknowledgement Top


The author is thankful to Director and other Administrators for their valuable cooperation during this study.

 
 ¤ References Top

1.Kelkar SS, Khare PM. Gel immune diffusion Techniques in Research and Laboratory Medicine. P 19, 154­-155 Popular Prakashan, Bombay, 1984.  Back to cited text no. 1      
2.Grunbaum BW, Lyons MF, Caroll N, Zee J. Quantitative Analysis of Normal Human Serum Proteins on Permanently Transpareatized Cellu­lose Acetate membranes. Micro­ chem J, 1963;7:54.  Back to cited text no. 2      
3.Kohn J, Feinberg JG. Electropho­ resis on Cellulose Acetate. Shan­den Instruments Co. Application Bull. 11, London 1965.  Back to cited text no. 3      
4.Smithies DJ. Zone Electrophoresis in starch gels and its application'to studies of serum proteins. Adv Protein Chem 1958:14:65.  Back to cited text no. 4      



This article has been cited by
1 The effect of different membrane buffers on hemoglobin electrophoresis
Liu, Z.-J., Jiang, H.-H.
Laboratory Hematology. 2001; 7(2): 101-102
[Pubmed]



 

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