| Condition |
Lysozyme |
Carbonic Anhydrase |
RNAse A |
BSA |
Myoglobin |
| Peptide hits * |
% Sequence Coverage** |
Peptide hits |
% Sequence Coverage |
Peptide hits |
% Sequence Coverage |
Peptide hits |
% Sequence Coverage |
Peptide hits |
% Sequence Coverage |
| Native protein mixture |
67/10 |
66% |
25/10 |
56% |
10/6 |
63% |
34/23 |
54% |
6/5 |
38% |
| FFPE; pH 4, 40 Kpsi |
26/8 |
69% |
9/7 |
36% |
12/5 |
59% |
21/12 |
26% |
3/3 |
28% |
| FFPE; pH 4, 14.7 psi |
4/1 |
15% |
n.d |
|
1/1 |
7% |
n.d |
|
n.d |
|
| FFPE; pH 8, 40 Kpsi |
75/7 |
57% |
12/7 |
30% |
11/5 |
71% |
23/15 |
29% |
3/2 |
16% |
| FFPE; pH 8, 14.7 psi |
n.d |
|
n.d |
|
n.d |
|
n.d |
|
n.d |
|
Multi-protein FFPE tissue surrogates were extracted at 100°C for 30 min followed by 80°C for 2 h at either 40,000 psi or atmospheric pressure (14.7 psi) in 50 mM Tris-
HCL, 2% (w/v) SDS buffer, pH 4 or 8. The extracts were washed extensively, and digested overnight with trypsin at 37°C in 50 mM NH4HCO3, pH 7.9 with 20% acetonitrile
(v/v). * Peptide hits: total spectra/number of unique peptides. ** % Sequence coverage: percent of theoretical tryptic peptides identified by LC/MS/MS. n.d.-none detected. |
