International Journal of Pure & Applied Bioscience (IJPAB)
Year : 2017, Volume : 5, Issue : 6
First page : (1) Last page : (8)
Article doi: http://dx.doi.org/10.18782/2320-7051.5988
Effect of Protein Extraction Buffer and Blocking Buffer on Efficiency of Prepared Nitrocellulose Membrane for Assembly of Lateral Flow Immunoassay Strip
Muhammad Irfan1*, Muhammad Asif1, Aftab Bashir1+ and Kauser Abdullah Malik1++
1*Gene Isolation Lab,
National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad
1+, 1++ Department of Biological Sciences, Forman Christian College, Lahore, Pakistan
*Corresponding Author E-mail: irfan.muhammad26@outlook.com
Received: 13.11.2017 | Revised: 5.12.2017 | Accepted: 8.12.2017
ABSTRACT
Lateral flow immunoassay strip is a simple, reliable and rapid qualitative tool for primary screening of transgenic proteins. The extraction of proteins from sample products for performing lateral flow immunoassay (LFIA) strip test is an important step. In this study, the protein extraction efficiency of various buffers prepared in-lab and some extraction buffers from commercial kits were tested. In addition, the effects of blocking buffers along with hybridization timings, for blocking of unoccupied sites, on prepared nitrocellulose membranes were examined during assembly of LFIA strips. It was estimated that 1XPBS of pH 7 is best protein extraction buffer according to our sample, Bt-Cry 1Ac from Bt-Cotton seeds. Furthermore, during manual assembly of LFIA strips, we found that the sealing tapes used as covering for one-unit assay has no effect on efficiency of LFIA strip tests. In our experiments, we optimized hybridization timing for blocking of unoccupied sites on nitrocellulose membranes is 30 minutes. We also optimized a simple composition for blocking buffer.
Key words: Transgenic proteins, Lateral flow immunoassay, Nitrocellulose membranes, Bt-Cotton, Protein extraction buffer
Full Text : PDF; Journal doi : http://dx.doi.org/10.18782
Cite this article: Irfan, M., Asif, M., Bashir, A., Malik, K.A., Effect of Protein Extraction Buffer and Blocking Buffer on Efficiency of Prepared Nitrocellulose Membrane for assembly of Lateral Flow Immunoassay Strip, Int. J. Pure App. Biosci.5(6): 1-8 (2017). doi: http://dx.doi.org/10.18782/2320-7051.5988
