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types of protein testing

 

LC-MS/MSLiquid Chromatography-Mass Spectrometry (LC-MS/MS) is an analytical process widely used in proteomics because it combines the physical separation capabilities of liquid chromatography with the mass determination ability of mass spectrometry in one integrated workflow.  LC-MS/MS enables the researcher to simultaneously identify proteins and post translational modifications, as well as perform protein quantitation using a label or label free techniques. LC-MS/MS can be used to analyze simple and complex biological samples.

 

2D-DIGETwo Dimensional – Difference in Gel Electrophoresis (2D-DIGE) is the most accurate, reproducible, and flexible gel-based approach for comparative and quantitative proteomics using distinctive fluorescent tags. With 2D-DIGE, 2 – 3 samples can be analyzed on one gel, and with a universal internal standard, multiple gel can be run and analyzed at the same time. 2D-DIGE can be applied to protein expression profiling as well as comparative proteomics using 2D analysis software to identify the proteins with a statistically significant difference in expression. 2D-DIGE provides a wide range of opportunities for researchers in Cardiovascular diseases, Cancer and other related fields; the 2D-DIGE analysis can uncover important proteins associated with these conditions

 

MudPITMultidimensional Protein Identification Technology (aka, MudPIT) is a liquid chromatography based proteomic technique used for separating and identifying complex proteins and peptides when the interest is to identify all the proteins in the biological sample is analyzed. The peptide mixture is prepared and pre-fractionated on or offline using strong cation exchange followed by online reverse phase LC-MS/MS for peptide and protein identification.  The data is then searched against a species-specific database for protein identification.

 

iTRAQ Isobaric tags for relative and absolute quantitation (iTRAQ) is a proteomic technique that utilizes multiplexed isobaric chemical tags which allows multiplexing of up to eight biological samples followed by LC-MS/MS.   The iTRAQ reagents consist of an amine reactive group, a reporter group, and a balance group. Simultaneous protein identification and quantitation are possible when the iTRAQ labeled peptides are analyzed by LC-MS/MS.  The labeling preserves post-translational modifications (PTMs) allowing for PTM analysis as well.

 

TMT Tandem mass tags, similar to iTRAQ, allow simultaneous protein identification and quantitation but have a higher plexing power.  Using TMT allows a researcher to multiplex up to 16 biological samples.

 

Luminex xMAP Luminex xMAP (Multi-Analyte Profiling) technology is designed to simultaneously detect multiple targeted proteins including cancer markers cytokines, growth factors chemokines, etc.., in the same sample. This technology combines advanced fluidics, optics, and digital signal processing with proprietary microsphere technology to deliver multiplexed assay capabilities. Featuring a flexible, open-architecture design, xMAP Technology can be configured to perform a wide variety of protein or nucleic acid assays quickly, cost-effectively, and accurately. xMAP Technology enables multiplexing of biological tests and produces results that are more efficient and comparable to traditional methods such as ELISA, western blotting, PCR, and traditional arrays.  The Luminex xMAP is an open source platform so there are many commercially available kits available or a custom analyte panel can be developed.

 
The choice of your protein identification method depends on your sample type, project goals, and budget…. At ITSI-Biosciences, we pride ourselves in the provision of standard and state-of-the-art protein testing services and have a wide array of bioanalytical kits for your proteomic and protein testing projects.

ITSI-Biosciences offers comprehensive bio-analytical solutions, accurate results done at a fast turnaround time. Contact us today for a quote and make your protein testing and analysis stress free. You can reach us via Email; itsi@itsibio.com, Phone; 814-262-7331, or Fax; 814-262-7334.