Bottom-up proteomic analysis of single HCT 116 colon carcinoma multicellular spheroids.

ABSTRACT:

Rationale

Proteomic analysis of single multicellular spheroids has not been previously reported. As three-dimensional cell cultures are an increasingly popular model system for biological research, there is interest in obtaining proteomic profiles of these samples. We investigated the proteome of single HCT 116 multicellular spheroids using protocols optimized for small sample sizes.

Methods

Six biological replicates were analyzed via microscopy for size. Total protein content was assessed via the bicinchoninic acid assay (BCA assay). Five separate biological replicate spheroids were analyzed via mass spectrometry in technical duplicate. An ultra-performance liquid chromatography (UPLC) system coupled with an LTQ Orbitrap Velos was used for peptide separation, analysis, and

SUBMITTER: Feist PE 

PROVIDER: S-EPMC4763982 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature