Detection of Isoforms Differing by a Single Charge Unit in Individual Cells.
To measure protein isoforms in individual mammalian cells, we report single-cell resolution isoelectric focusing (scIEF) and high-selectivity immunoprobing. Microfluidic design and photoactivatable materials establish the tunable pH gradients required by IEF and precisely control the transport and handling of each 17-pL cell lysate during analysis. The scIEF assay resolves protein isoforms with resolution down to a single-charge unit, including both endogenous cytoplasmic and nuclear proteins from individual mammalian cells.
The UC Berkeley/UCSF Graduate Program in Bioengineering, Berkeley, CA, USA.|Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Augusto M
0000-0003-4596-0059
AM
Tentori
The UC Berkeley/UCSF Graduate Program in Bioengineering, Berkeley, CA, USA.
Kevin A
0000-0002-7818-1388
KA
Yamauchi
The UC Berkeley/UCSF Graduate Program in Bioengineering, Berkeley, CA, USA. aeh@berkeley.edu.|Department of Bioengineering, UC Berkeley, 308B Stanley Hall, Berkeley, CA, 94720, USA. aeh@berkeley.edu.
Amy E
0000-0002-6906-2985
AE
Herr
D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007525:Isoelectric Focusing / Q000379:methods*; D056656:Lab-On-A-Chip Devices; D020033:Protein Isoforms / Q000032:analysis*; D059010:Single-Cell Analysis
D005456:Fluorescent Dyes; D020033:Protein Isoforms
immunoassays; isoelectric focusing; lab-on-a-chip; proteomics; single-cell analysis