A new technique for the isolation and purification of the basal lamina from insect tissues.
A new technique for the isolation and purification of basal lamina from insect tissues using cell dissociation at pH 2 is described. Tissue incubation in these solutions results in the spontaneous detachment of cells from the basal lamina which can be collected free of any significant contamination by cellular components. Short lengths of plasma membrane which remain attached to the basal lamina can be removed by subsequent sonication or detergent treatment. Using Malpighian tubules as the primary test tissue, we have found that the procedure only requires a few minutes, works equally well on pooled tissue samples, individual tissue pieces or tissue subregions and involves no loss of basal lamina from the starting material.
D000818:Animals; D001485:Basement Membrane / Q000648:ultrastructure; D002458:Cell Fractionation / Q000379:methods*; D002951:Citrates; D019343:Citric Acid; D006863:Hydrogen-Ion Concentration; D007915:Lepidoptera / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D013010:Sonication; D013501:Surface-Active Agents
D002951:Citrates; D013501:Surface-Active Agents; D019343:Citric Acid