Information for Peptide: FPELNYQLK from STAT3_HUMAN

1. Biological Peptide

1.1 Native Sequence

    K.FPELNYQLK.I

1.2 Peptide Properties

Start	Stop	MW	m/z for 2+	m/z for 3+	pI
355	363	1150.6	576.31	384.54	6.0

1.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
F	b		y
P	b		y
E	b		y
L	b		y
N	b		y
Y	b		y
Q	b		y
L	b		y
K	b		y

2. Synthetic Peptide

2.1 Synthetic (Internal Standard) Peptide Sequence

FPELNYQL(C6N1)K (Modification: Stavle Isotope on L8 (U-13C6, 15N) )

2.2 Peptide Properties

MW	m/z for 2+	m/z for 3+	pI
1157.62	579.82	386.88	6.0

2.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
F	b		y
P	b		y
E	b		y
L	b		y
N	b		y
Y	b		y
Q	b		y
L	b		y
K	b		y

2.4 Peptide Synthesis Report

Click to view/download synthesis report for FPELNYQLK

2.5 QqQ MS/MS

Transition Table:

Transition	m/z of Transition	Collision Energy
y5	672.36229	23.0
y6	785.44636	23.0
y7	914.48895	23.0
y8	1011.54171	23.0

2.6 Calibration Curve of Standard

Description: NONE

 

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