Membrane Proteomics

Membrane Proteomics or Membrane Protein Array with 2D DIGE focuses on detecting and quantifying membrane protein complex and their post translational modifications (PTMs), with high resolution and accuracy. We also offer nanoLC-MS/MS to study membrane proteomics.

Membrane proteins normally have higher pI and contain various PTMs. They are much harder to extract due to a higher insolubility. In general, they show poor resolution on standard 2D gels.

Applied Biomics has developed unique methods to efficiently extract membrane proteins from the prepared membrane fractions. In addition, we have optimized IEF and 2D electrophoresis conditions to achieve an optimal resolution of the membrane proteins.

For gel layout, please view the study designs based on the total number of samples.

Example 1:

The figure below shows 2D DIGE analysis of the membrane proteins from S phase and G2/M phase. The membrane fractions were prepared from HT29 cells using sucrose density gradient. Membrane proteins were extracted from membrane fractions using Applied Biomics’ membrane protein extraction protocol. The 2D DIGE gel clearly revealed some significantly changed membrane proteins between S and G2/M phase with high 2D gel resolution.

S-Phase Membrane Protein

Membrane proteomics: Black/white image of S-phase membrane proteome

G2/M-Phase Membrane Protein

Membrane proteomics: Black/white image of G2/M-phase membrane proteome

S-Phase / G2/M-Phase

Membrane proteomics: overlay image of S- and G2/M-phase membrane proteome

S-Phase Membrane Protein

G2/M-Phase Membrane Protein

S-Phase / G2/M-Phase

Membrane proteomics: overlay image of S- and G2/M-phase membrane proteome

Sample Info

Please follow these general principles regarding biohazardous materials.

Sample amount: 150-250 µg of membrane fractions from each sample. This will be sufficient to cover the entire 2D DIGE procedure including analytical gel, preparative gel, spot picking and protein ID.

Protein concentration: 5-20 mg/mL

Buffer: Please feel free to submit samples in whichever buffer that you would normally use. The standard sample preparation is covered by the DIGE gel cost. Such samples should have protein concentration in the range of 5-20 mg/ml. Additional charge may apply on samples that require extensive amount of the extra work such as protein eluting, concentrating and buffer exchange.

Pricing

Please view here for Membrane Proteomics pricing.