Western Blot Analysis of Cells Encapsulated in Self-Assembling Peptide Hydrogels

The continuous optimisation of current analytical techniques in vitro is ever more pressing with the development of new materials for applications in tissue engineering. In particular, the isolation of cellular components for downstream applications is often hindered by the presence of biomaterials and a major obstacle in understanding how cell-matrix interactions influence cell behaviour. We present an approach to carryout western-blot analysis on cells encapsulated in self-assembling peptide hydrogels (SAPH) which highlights the need for complete solubilisation of the hydrogel construct. We demonstrate that both the choice of buffer and multiple cycles of sonication are vital in obtaining complete solubilisation, thereby enabling the detection of proteins otherwise lost to SAP aggregation. Moreover, we show that the presence of SAP does not interfere with standard immunoblotting technique, offering potential for use in more full-scale proteomic studies.