Development of an antibody against a 170-kDa fragment of fibroin isolated from cocoon fibres of Bombyx mori.
A simple method for the isolation of denatured fragments of the fibroin protein from cocoon fibres by alkali solubilization is discussed. This 170-kDa antigen has been purified and used to raise the polyclonal antibody in rabbit. The specificity of this antibody to the purified cocoon protein has suggested strong immunoreactivity up to a titre of 1:5000 dilution of the antibody. Further, dot-blot analysis with the tissue extracts from silk glands of different larval stages (3rd to 5th) reveals that this antibody reacts showing a stage-specific increase in the intensity of the colour, correlating well with the in-vivo expression of the silk protein. This study suggests the availability of a specific polyclonal antibody that detects the native fibroin with no crossreactivity with other tissue proteins.
Rajan
M K
MK
Centre for Biotechnology, A.C. College of Technology, Anna University, Madras, India.
Balakrishnan
A
A
Jayaraman
K
K
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D012831:Bombyx / Q000737:chemistry* / Q000502:physiology; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005349:Fibroins / Q000276:immunology / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007814:Larva; D008970:Molecular Weight; D010446:Peptide Fragments / Q000276:immunology / Q000302:isolation & purification*; D012394:Rosaniline Dyes; D012995:Solubility; D014020:Tissue Extracts / Q000032:analysis
D010446:Peptide Fragments; D012394:Rosaniline Dyes; D014020:Tissue Extracts; C048139:Coomassie blue; D005349:Fibroins