Entering edit mode
10.1 years ago
jgbradley1
▴
110
I'm new to Bioinformatics and I'm trying to learn how to quantify peptides from an iTRAQ experiment. I know with SILAC, everyone uses the log(H/L) ratio because it reflects the relative protein abundance. How do I calculate the relative protein abundance in an iTRAQ experiment (assuming 4-plex)? I'm hoping someone can provide a worked-out example so I can understand what's happening better. I've pulled one row of data from an iTRAQ experiment file. If you need additional explanation of each field in the table, please see this file. It looks like the following:
QueryPrecursorMz 400.5555
OriginalPrecursorMz 400.5555
PrecursorError(ppm) 0.45712903
QueryCharge 3
OriginalCharge 3
PrecursorScanNum 1
PrecursorArea 9.55E+08
PrecursorRelAb 0.00012481
RTAtPrecursorHalfElution 45
PeptideSequence +144.102HNQRPTFR
AmbiguousMatch 0
Protein gi|160420317|ref|NP_001104026.1|(pre=K,post=Q);gi|116063573|ref|NP_001447.2|(pre=K,post=Q)
DeNovoScore 68
MSGFScore 68
Evalue 8.58E-05
Qvalue 8.48E-05
PepQvalue 1.49E-04
PrecursorPurity 63.4,72.8
FractionDecomposition 94.7,94.7
HCDEnergy 19.0664520263672eV
iTRAQ114 16563.3/0.09
iTRAQ115 118556/0.10
iTRAQ116 157171/0.09
iTRAQ117 119378/0.09
iTRAQFlags I
iTRAQTotalAb 411668
iTRAQFractionOfTotalAb 0.136185
I have seen A: On Calculating I-Traq Ratio previous post, however the link to the worked-out example is dead.