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.
Applied Biomics has developed unique methods to efficiently extract membrane proteins from prepared membrane fractions, overcoming challenges associated with their higher pI and low solubility. In addition, we have optimized IEF and 2D electrophoresis conditions to achieve optimal resolution of membrane proteins.
Membrane Proteomics Applications
- Suitable sample types: Membrane fractions
- Drug target discovery
- Cancer research
- Cell signaling
- Cell-surface protein profiling
- Characterization of receptors, transporters, & ion channels
Membrane Proteomics Workflow
- Membrane protein extraction from membrane fractions
- Fluorescent sample labeling
- 2D gel electrophoresis
- Image acquisition & image analysis
- Quantitative analysis
- Automated spot picking
- LC-MS/MS protein identification
- Pathway analysis & validation analysis (optional)
Membrane Proteomics Deliverables
- Image report
- High-resolution 2D proteome images for each sample
- Overlay images comparing two samples
- Mark differentially expressed spots
- Quantitation report
- Protein abundance ratios for differentially expressed spots
- Statistical P-value (if applicable)
- Protein ID report
- Protein Level: Protein identity, abundance, sequence coverage, and confidence metrics
- Peptide Level: Matched peptide sequences, masses, and confidence metrics
Membrane Proteomics Study Design & Exmaples
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
G2/M-Phase Membrane Protein
S-Phase / G2/M-Phase
S-Phase Membrane Protein
G2/M-Phase Membrane Protein
S-Phase / G2/M-Phase
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 sample preparation including protein extraction (if applicable), sample cleaning and measuring protein concentration is not covered by the 2D DIGE gel cost. Additional charge may apply on samples with concentration less than 4 mg/mL, or require significant amount of the extra work such as protein eluting, sample pooling, extensive buffer exchange or serum abundant protein depletion.
Sample type: Please inform our scientists about your sample type. We will send you additional tips for each sample type.
Pricing
Please view here for Membrane Proteomics pricing.