Information for Peptide: APVSGTQEK from RAF1_HUMAN

1. Biological Peptide

1.1 Native Sequence

    R.APVSGTQEK.N

1.2 Peptide Properties

Start	Stop	MW	m/z for 2+	m/z for 3+	pI
319	327	915.47	458.74	306.16	6.05

1.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
A	b		y
P	b		y
V	b		y
S	b		y
G	b		y
T	b		y
Q	b		y
E	b		y
K	b		y

2. Synthetic Peptide

2.1 Synthetic (Internal Standard) Peptide Sequence

APVSGTQDK (Modification: Amino acid switch was made from E8 to D )

2.2 Peptide Properties

MW	m/z for 2+	m/z for 3+	pI
901.45	451.73	301.49	5.88

2.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
A	b		y
P	b		y
V	b		y
S	b		y
G	b		y
T	b		y
Q	b		y
D	b		y
K	b		y

2.4 Peptide Synthesis Report

Click to view/download synthesis report for APVSGTQEK

2.5 QqQ MS/MS

Transition Table:

Transition	m/z of Transition	Collision Energy
y8	831.42126	18.0
y7	734.3685	19.0
y6	635.30009	20.0
y5	548.26806	18.0
y4	491.2466	20.0
y3	390.19892	23.0
b4	355.19819	17.0
b3	268.16616	18.0

2.6 Calibration Curve of Standard

Description: A dilution series was done on the internal standard peptide from 50 Fmol to 0.1 fmol. Peptide detection was possible to 5 fmol.

 

Experiment Description: Cells from Melanoma cell lines were lysed in 8M urea/ 100mM ammonium bicarbonate buffer on ice. The equivalent of 200,000 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. 1/6 of the resulting digest was then analyzed on a Thermo Scientific TSQ mass spectrometer.

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