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Why does serum/plasma proteomics work the best with 2D DIGE but not LC-MS/MS?
Because 2D DIGE can generate much better serum proteomics data at a lower cost. Analysis of serum and plasma proteome represents a challenge due to the wide range of protein concentrations and high structural complexity of the constituent proteins in these samples. In serum and plasma, the quantities of high- and some low-abundance proteins span over 10 orders of magnitude. Immunodepletion has been used to enhance the detection of low abundance plasma proteins. However, researches have discovered that, even with immunodepletion, low abundance proteins still remain undetectable using LC-MS/MS platform. Furthermore, immunodepletion process will: (i) remove proteins associated with the abudant proteins, (ii) cause protein degradations, and (iii) introduce variations, thus compromising the data quality.
Applied Biomics developed a Serum and Plasma 2D DIGE proteomics protocol by which thousands of low-abundance serum proteins can be resolved and quantitatively analyzed WITHOUT immunoaffinity depletion of the serum abundant proteins. This technology allows for a more sensitive detection and accurate quantification of low-abundance serum/plasma proteins. For details please view see our Publication on Genetic Engineering & Technology News. Such platform has been successfully applied in discovering disease- or treatment-specific serum/plasma biomarkers. We offer the following services in serum/plasma proteomics:
A. Improved Serum 2D DIGE Without Immuno-Depletion
B. Improved Serum 2D DIGE Combined With Immuno-depletion Of Abundant Proteins
A. Improved Serum 2D DIGE Without Immuno-depletion
* Example 1: Human Serum Sample. The left, middle and right image shows the human serum proteome using standard protocol, Applied Biomics (ABO) improved protocol with pH3-10, and pH4-7, respectively. The ABO protocol clearly reveals much more low aboundant serum proteins than the standard protocol.
Standard 2D Protocol
(pH3-10)
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ABO pH3-10 Protocol
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ABO pH4-7 Protocol  |
* Example 2: Preclinical Trial Mouse Serum Before and After Drug Feeding Using ABO Protocol. Four pairs of mouse serum samples were collected before and after 4 different treatments, then analyzed by using Applied Bimics' improved serum protocol. The overlay images showed the difference between the serum samples collected before (green color) and after (red color) the drug treatment. The circled spots (in red) indicated a protein with increased abundance due to drug treatment. The 3-D view of the same spot is displayed in the right panel.
B. Improved Serum 2D DIGE Combined With Immuno-depletion Of Abundant Proteins
The Challenge of Biomarker Discovery from Plasma/Serum
Transferrin: 76 kDa
Fibrinogen: 65 kDa (a), 51 kDa (g) a1-antitrypsin: 56 kDa IgG: 150 kDa HSA: 67 kDa IgA: 180 kDa IgM: 750 kDa Transferrin: 79 kDa a2-Macroglobulin: 160 kDa Haptoglobin: 43 kDa alpha 1-acid glycoprotein: 23 kDa APO A-I, APO A-II: 23 kDa |
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| Protein dynamic range: 10 magnitude of difference in amount |
We offer the following serum abundant protein immuno-depletion services:
1) Human Serum: Albumin/IgG depletion
2) Human Serum: IgY-12 depletion (12 most abundant human plasma proteins: Human Serum Albumin, IgG, Fibrinogen, Transferrin, IgA, IgM, Haptoglobin, a2-Macroglobulin, a1-Acid Glycoprotein, a1-Antitrypsin and HDL (Apo A-I & Apo A-II).
3) Mouse Serum: Albumin/IgG depletion
4) Rat Serum: Albumin/IgG depletion
5) Monkey Serum: Albumin/IgG depletion
* Example: Human plasma with and without IgY-12 Immuno-depletion (pH4-7)
Plasma Without Immuno-Depletion
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Plasma With Immuno-Depletion
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If you are interested, please follow these steps to obtain more information and order this service.