Home > Proteomics Services > iTRAQ Service
We offer iTRAQ service, a liquid-based proteomics analysis. It labels the Lysine residues and N terminus of all peptides with isotopes of identical masses, thus allows accurate quantitative comparisons between 2 (2-plex) to 8 (8-plex) samples. This approach can provide both protein identification and protein ratios in one experiment. In our iTRAQ service, we have made the following improvements that have significantly increased the sensitivity of detecting low abundant proteins and the total number of proteins (see an example here):
1) Optimized protein extraction on samples such as serum, clinical samples with interfering substances, protein complex and whole cell lysate
2) Optimized a proprietary fractionation protocol specifically targeting the low-abundant basic proteins (pI > 8.0), such as membrane proteins (see example)
3) Extended nanoLC gradient coupled with optimized MSMS parameters, totaling a 12 hour instrument time
We have optimized protein extraction for iTRAQ analysis on the following sample types:
1) Whole cell lysate from cultured cells, bacteria and other organisms
2) Subcellular organelles such as Mitochondria, Lysosome and Exosome etc
3) Protein complex such as IP or column purified complex with very low protein amount (<2ug)
4) Clinical samples with interfering substances such as high salt in urine
5) Serum samples with high abundant proteins and a wide dynamic range
Protein samples |
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Reduction/Alkylation & Trypsin Digestion |
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iTRAQ reagent labeling |
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Mix and clean up samples |
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NanoLC-MS/MS |
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MS |
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MS/MS |
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Reporter ion |
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Example-1: Characteristics Of Proteins Detected In A Single iTRAQ Experiment
Protein Ratios (0.14 to 3.7)![]() |
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Protein Mass (8.5 to 800KD)![]() |
Protein pI (3.7 to 12) |
Example-2: Characteristics Of Proteins Detected In An iTRAQ Experiment Using High pI Fractionation
Protein Ratios (0.46 to 3.1)![]() |
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Protein Mass (8.5 to 622KD)![]() |
Protein pI (4.6 to 11.4) |
* 257 out of 545 proteins (~50%) of identified proteins are basic (pH >=8.0).
Top 25 (Out Of 545) Hits Obtained In High pI Fractionation iTRAQ Experiment
Rank | Ratios | MW (Da) | pI | Accession | Protein Name |
---|---|---|---|---|---|
1 |
0.92 |
120,027 |
7.5 |
MTP_MOUSE |
Microsomal triglyceride transfer protein large subunit OS=Mus musculus GN=Mttp PE=2 SV=2 |
2 |
0.88 |
43,498 |
8.9 |
MDHM_MOUSE |
Malate dehydrogenase, mitochondrial OS=Mus musculus GN=Mdh2 PE=1 SV=3 |
3 |
0.91 |
49,409 |
8.3 |
THIM_MOUSE |
3-ketoacyl-CoA thiolase, mitochondrial OS=Mus musculus GN=Acaa2 PE=1 SV=3 |
4 |
0.83 |
69,450 |
9.2 |
ATPA_MOUSE |
ATP synthase subunit alpha, mitochondrial OS=Mus musculus GN=Atp5a1 PE=1 SV=1 |
5 |
0.85 |
44,001 |
8.4 |
G3P_MOUSE |
Glyceraldehyde-3-phosphate dehydrogenase OS=Mus musculus GN=Gapdh PE=1 SV=2 |
6 |
0.87 |
46,479 |
8.5 |
ALDOB_MOUSE |
Fructose-bisphosphate aldolase B OS=Mus musculus GN=Aldob PE=1 SV=3 |
7 |
0.81 |
40,241 |
8.6 |
VDAC1_MOUSE |
Voltage-dependent anion-selective channel protein 1 OS=Mus musculus GN=Vdac1 PE=1 SV=3 |
8 |
0.96 |
56,723 |
9.3 |
QCR2_MOUSE |
Cytochrome b-c1 complex subunit 2, mitochondrial OS=Mus musculus GN=Uqcrc2 PE=1 SV=1 |
9 |
1.23 |
69,368 |
7.2 |
KPYM_MOUSE |
Pyruvate kinase isozymes M1/M2 OS=Mus musculus GN=Pkm2 PE=1 SV=4 |
10 |
1.17 |
48,995 |
7.6 |
ANXA2_MOUSE |
Annexin A2 OS=Mus musculus GN=Anxa2 PE=1 SV=2 |
11 |
1.18 |
40,911 |
9.1 |
LEG4_MOUSE |
Galectin-4 OS=Mus musculus GN=Lgals4 PE=1 SV=2 |
12 |
1.85 |
66,905 |
7.9 |
AL1A1_MOUSE |
Retinal dehydrogenase 1 OS=Mus musculus GN=Aldh1a1 PE=1 SV=5 |
13 |
0.84 |
195,543 |
6.5 |
CPSM_MOUSE |
Carbamoyl-phosphate synthase [ammonia], mitochondrial OS=Mus musculus GN=Cps1 PE=1 SV=2 |
14 |
1.21 |
19,712 |
8.6 |
FABPL_MOUSE |
Fatty acid-binding protein, liver OS=Mus musculus GN=Fabp1 PE=1 SV=2 |
15 |
1.17 |
57,299 |
8.0 |
PGK1_MOUSE |
Phosphoglycerate kinase 1 OS=Mus musculus GN=Pgk1 PE=1 SV=4 |
16 |
0.97 |
17,990 |
8.5 |
PROF1_MOUSE |
Profilin-1 OS=Mus musculus GN=Pfn1 PE=1 SV=2
PE=1 SV=2 |
17 |
1.02 |
102,141 |
8.1 |
ACON_MOUSE |
Aconitate hydratase, mitochondrial OS=Mus musculus GN=Aco2 PE=1 SV=1 |
18 |
0.89 |
45,575 |
9.1 |
BDH_MOUSE |
D-beta-hydroxybutyrate dehydrogenase, mitochondrial OS=Mus musculus GN=Bdh1 PE=1 SV=2 |
19 |
1.20 |
110.361 |
5.7 |
VILI_MOUSE |
Villin-1 OS=Mus musculus GN=Vil1 PE=1 SV=3 |
20 |
1.34 |
87,074 |
8.6 |
ACOX1_MOUSE |
Peroxisomal acyl-coenzyme A oxidase 1 OS=Mus musculus GN=Acox1 PE=1 SV=5 |
21 |
0.97 |
49,097 |
8.7 |
THIKA_MOUSE |
3-ketoacyl-CoA thiolase A, peroxisomal OS=Mus musculus GN=Acaa1a PE=2 SV=1 |
22 |
0.95 |
64,684 |
9.1 |
EF1A1_MOUSE |
Elongation factor 1-alpha 1 OS=Mus musculus GN=Eef1a1 PE=1 SV=3 |
23 |
0.53 |
30,707 |
8.6 |
CELA1_MOUSE |
Chymotrypsin-like elastase family member 1 OS=Mus musculus GN=Cela1 PE=2 SV=1 |
24 |
1.01 |
41,646 |
8.9 |
DEOC_MOUSE |
Putative deoxyribose-phosphate aldolase OS=Mus musculus GN=Dera PE=2 SV=1 |
25 |
0.93 |
43,872 |
8.8 |
HCDH_MOUSE |
Hydroxyacyl-coenzyme A dehydrogenase, mitochondrial OS=Mus musculus GN=Hadh PE=1 SV=2 |
If you are interested, please follow these steps to obtain more information and order this service.
* iTRAQ is the trademark of AB SCIEX..