Information for Peptide: FAGVFHVEK from CD44_HUMAN

1. Biological Peptide

1.1 Native Sequence

    R.FAGVFHVEK.N

1.2 Peptide Properties

Start	Stop	MW	m/z for 2+	m/z for 3+	pI
30	38	1032.54	517.28	345.19	6.75

1.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
F	b		y
A	b		y
G	b		y
V	b		y
F	b		y
H	b		y
V	b		y
E	b		y
K	b		y

2. Synthetic Peptide

2.1 Synthetic (Internal Standard) Peptide Sequence

FAGVFHV(C5N1)EK (Modification: Stable Isotope on V7 (U-13C5, 15N) )

2.2 Peptide Properties

MW	m/z for 2+	m/z for 3+	pI
1038.55	520.28	347.19	6.75

2.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
F	b		y
A	b		y
G	b		y
V	b		y
F	b		y
H	b		y
V	b		y
E	b		y
K	b		y

2.4 Peptide Synthesis Report

Click to view/download synthesis report for FAGVFHVEK

2.5 QqQ MS/MS

Transition Table:

Transition	m/z of Transition	Collision Energy
y8	892.49	17.0
y7	821.46	20.0
y6	764.43	15.0
y5	665.37	21.0
y4	518.3	24.0
y3	381.24	22.0

2.6 Calibration Curve of Standard

Description: Internal standard FAGVFHVEK was diluted from 50 fmol to 500 amol injection concentrations and was found to be sensitive down to 5 fmol

 

Experiment Description: Cells from colon caner line HCT 116 were lysed in 8M urea/ 100mM ammonium bicarbonate buffer on ice. The equivalent of 100,000 cells were loaded onto a SDS gel. The protein of interest was excised and ingel digestion with trypsin was performed after reduction and alkylation with TCEP and IAA. 1/4 of the resulting digest was then analyzed on a Thermo Scientific TSQ mass spectrometer. Sample was analyzed concurrently with internal standard

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