Information for Peptide: ILHTAWHPK from 2ABA_HUMAN

1. Biological Peptide

1.1 Native Sequence

    K.ILHTAWHPK.E

1.2 Peptide Properties

Start	Stop	MW	m/z for 2+	m/z for 3+	pI
419	427	1101.61	551.81	368.21	8.76

1.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
I	b		y
L	b		y
H	b		y
T	b		y
A	b		y
W	b		y
H	b		y
P	b		y
K	b		y

2. Synthetic Peptide

2.1 Synthetic (Internal Standard) Peptide Sequence

ILHTAWHP(c5N1)K (Modification: Stable Isotope on P8 (u-13C5, 15N) )

2.2 Peptide Properties

MW	m/z for 2+	m/z for 3+	pI
1107.62	554.82	370.22	8.76

2.3 Fragment Ion Table

AA	b Ion Series	b Ion Mass	y Ion Series	y Ion Mass
I	b		y
L	b		y
H	b		y
T	b		y
A	b		y
W	b		y
H	b		y
P	b		y
K	b		y

2.4 Peptide Synthesis Report

Click to view/download synthesis report for ILHTAWHPK

2.5 QqQ MS/MS

Transition Table:

Transition	m/z of Transition	Collision Energy
y3	387.23	26.0
y4	573.3	18.0
y5	644.34	21.0
y6	745.39	22.0
y7	882.45	21.0
y8	995.53	22.0

2.6 Calibration Curve of Standard

Description: A dilution series was doneon the internal standard from 50 fmol to 0.1 fmol. The detection limit was determined to be 10 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