Information for Peptide: TLTDEELADWK from STAT3_HUMAN
1. Biological Peptide
1.1 Native Sequence
K.TLTDEELADWK.R
1.2 Peptide Properties
| Start | Stop | MW | m/z for 2+ | m/z for 3+ | pI |
|---|---|---|---|---|---|
| 234 | 244 | 1319.62 | 660.82 | 440.88 | 3.92 |
1.3 Fragment Ion Table
| AA | b Ion Series | b Ion Mass | y Ion Series | y Ion Mass |
|---|---|---|---|---|
| T | b | y | ||
| L | b | y | ||
| T | b | y | ||
| D | b | y | ||
| E | b | y | ||
| E | b | y | ||
| L | b | y | ||
| A | b | y | ||
| D | b | y | ||
| W | b | y | ||
| K | b | y |
2. Synthetic Peptide
2.1 Synthetic (Internal Standard) Peptide Sequence
TL(C6N1)TDEELADWK (Modification: Stable Isotope on L2 (U-13C6, N15) )
2.2 Peptide Properties
| MW | m/z for 2+ | m/z for 3+ | pI |
|---|---|---|---|
| 1326.64 | 664.33 | 443.22 | 3.92 |
2.3 Fragment Ion Table
| AA | b Ion Series | b Ion Mass | y Ion Series | y Ion Mass |
|---|---|---|---|---|
| T | b | y | ||
| L | b | y | ||
| T | b | y | ||
| D | b | y | ||
| E | b | y | ||
| E | b | y | ||
| L | b | y | ||
| A | b | y | ||
| D | b | y | ||
| W | b | y | ||
| K | b | y |
2.4 Peptide Synthesis Report
Click to view/download synthesis report for TLTDEELADWK
2.5 QqQ MS/MS
Transition Table:
| Transition | m/z of Transition | Collision Energy |
|---|---|---|
| y7 | 890.426 | 26.0 |
| y8 | 1005.452 | 26.0 |
| y9 | 1106.5 | 26.0 |
| y10 | 1226.584 | 26.0 |
2.6 Calibration Curve of Standard
Description: Dilution series obtained by direct infusion of peptide concentrations ranging from 50 fmol per microliter to 0.1 fmol per microliter with a 60 second aquisition time over which the average intensity was determined. Peptide was able to be detected down to a concentration of 5 fmole.
Experiment Description: 1 million cells from cell line K526 were lysed in an immuno-precipitation buffer containing 20 mM Tris-HCL (pH 7.6), 150 mM NaCl, 1mN EDTA, 0.1% NP-40, 100ug/ml PMSF, and 1mM sodium vanadate. Cell lysates were vortexed on ice for 30 minutes and spun to clear. The Supernatant was then incubated with protein A, and then with ABL1 specific antibody that has already been bound to protein A. Samples were incubated overnight, washed and then the beads were boiled in gel loading buffer and run for 1 hour on an SDS page gel (12% Bis-Tris). Resulting gel bands were then excised, reduced, alkylated and digested overnight with trypsin. Internal standard was injected concurrently
2.7 LC-MRM Analysis of Biological and Standard Peptides Illustrates Elution Times
2.8 Pseudo MS/MS Comparison of Transition Patterns for Biological/Standard Peptides
