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PROTEIN IDENTIFICATION

We offer high sensitivity protein identification from gel spots or bands using the latest technologies in mass spectrometry. In each run, four sensitivity standards in the amount of 1 femtomole are included. Our sensitivity is 1 femtomole.

Sample type : well resolved 1D or 2D gel spots containing only 1 protein species

Guidelines for Preparing and Shipping Samples for Protein Identification

Whole Genome Resequencing
  • Database search : Protein identification is based on peptide fingerprint mass mapping (using MS spectra) and peptide fragmentation mapping (using MS/MS spectra). Combined MS and MS/MS spectra are submitted for database search using GPS Explorer software equipped with the MASCOT search engine to identify proteins from primary sequence databases.
  • Gel treatment : 2D gel spots are washed multiple times to remove staining dye and other inhibitory chemicals. Gel spots are then dried to absorb maximum volume of digestion buffer.
  • Data Report : Includes the top 10 highest scoring hits from the database search for each 2D gel spot, as well as a summary listing the best match for each sample.
  • In-gel trypsin digestion : Dried 2D gel spots are rehydrated in digestion buffer containing sequencing grade modified trypsin. Proteins are digested in-gel at 37°C.
  • MALDI-TOF/TOF : Ten to twenty of the most abundant peptides in each sample are further subjected to fragmentation and tandem mass spectrometry (MS/MS) analysis.
  • Spotting : The desalted peptides are mixed with CHCA matrix (alpha-cyano-4-hydroxycinnamic acid) and spotted into wells of a MALDI plate.
  • MALDI-TOF : Mass spectra (MS) of the peptides in each sample are obtained using an Applied Biosystems Proteomics Analyzer.
  • Peptide extraction : Digested peptides are extracted from gel with TFA extraction buffer and shaking.
  • Desaltin: The digested tryptic peptides are desalted using C-18 Zip-tips (Millipore).

About Us

NGB Diagnostics Pvt. Ltd. founded in the year 2012, is an emerging Indian genomics company, with the mission of facilitating a wide array of Next generation genomic and proteomic services, high-quality molecular biology products, integral solutions for the clinical diagnostics, Pharma, R&D and Agro-food sectors.

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