|
|
ORIGINAL CONTRIBUTION |
|
|
|
| Year : 1997 | Volume
: 51
| Issue : 4 | Page : 113-114 |
| |
Reuse of cellulose acetate membrane strips for protein and haemoglobin electrophoretic analysis
MM Satwekar
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
PMID: 9355695
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 |
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 | |  |
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 | | |
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 | | |
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 | | |
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 | | |
The author is thankful to Director and other Administrators for their valuable cooperation during this study.
| ¤ References | | |
| 1. | Kelkar SS, Khare PM. Gel immune diffusion Techniques in Research and Laboratory Medicine. P 19, 154-155 Popular Prakashan, Bombay, 1984.  |
| 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. |
| 3. | Kohn J, Feinberg JG. Electropho resis on Cellulose Acetate. Shanden Instruments Co. Application Bull. 11, London 1965. |
| 4. | Smithies DJ. Zone Electrophoresis in starch gels and its application'to studies of serum proteins. Adv Protein Chem 1958:14:65. |
| 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] | |
|
|
|
|
