Information for Peptide: GEPWFHGK from SHC1_HUMAN

1. Biological Peptide

1.1 Native Sequence

    R.GEPWFHGK.L

1.2 Peptide Properties

Start	Stop	MW	m/z for 2+	m/z for 3+	pI
485	492	956.45	479.23	319.82	6.75

1.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
G	b		y
E	b		y
P	b		y
W	b		y
F	b		y
H	b		y
G	b		y
K	b		y

2. Synthetic Peptide

2.1 Synthetic (Internal Standard) Peptide Sequence

GEPWF(C9N1)HGK (Modification: Stable Isotope on F5 (U-13C9, N15) )

2.2 Peptide Properties

MW	m/z for 2+	m/z for 3+	pI
966.48	484.25	323.17	6.75

2.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
G	b		y
E	b		y
P	b		y
W	b		y
F	b		y
H	b		y
G	b		y
K	b		y

2.4 Peptide Synthesis Report

Click to view/download synthesis report for GEPWFHGK

2.5 QqQ MS/MS

Transition Table:

Transition	m/z of Transition	Collision Energy
y4	498.26219	20.0
y5	684.3415	20.0
y6	781.39426	20.0
y7	910.43685	20.0

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 0.1 fmole.

 

Experiment Description: Cells from Melanoma cell lines were lysed in RIPA buffer. The equivalent of 200,000 cells were loaded onto an SDS gel. The protein of interest was excised and ingel digestion with trypsin was performed after reduction and alkylation with TCEP and IAA. 1/6 of the resulting digest was then analyzed on a Thermo Scientific TSQ mass spectrometer. 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