PCR

TWO DIMENSIONAL GEL ELECTROPHORESIS SYSTEM


Working principles (With illustration)

Two-dimensional gel electrophoresis (or 2DE) is a powerful and widely used method for the analysis of complex protein mixtures extracted from cells, tissues, or other biological samples. This technique separate proteins in two steps: the first-dimension gel electrophoresis is isoelectric focusing (IEF), which separates proteins according to their isoelectric points (pI); the second-dimension is SDS-polyacrylamide gel electrophoresis (SDS-PAGE), which separates proteins according to their molecular weights (MW). In this way, complex mixtures consisted of thousands of different proteins can be resolved and the relative amount of each protein can be determined. Two-dimensional gel electrophoresis (2-DE) is an important tool for proteomics. The 2-DE can achieve the separation of several thousand different proteins in one gel. Differentially expressed proteins can be subsequently identified by mass spectrometric methods.

Application and studies related to

  • Comparative proteomics research.

    • User Instruction (how to bring samples)

  • The cells, tissues, or other biological samples (~1.0 gm in weight) or pure/crude protein samples should be in perfectly cold conditions (dry ice/liquid nitrogen/ice packs/ice). Avoid repeated change in storage conditions of sample(s). All samples should be freshly dissected/prepared in standard conditions.

    • User name:
      Praveen Maurye
    • Instrument Details:
    • Make
      Amersham Biosciences
    • Model:
      Ettan IPGphor 3 IEF system (for first dimension gel electrophoresis) & SE 600 vertical unit (for second dimension gel elctrophoresis).
    • Specification

      A fully integrated isoelectric focusing system optimized to deliver speed and reliability in the first dimension of 2DE. In our institute, we can run up to 12 IPG strips (7 or/& 11 cm) simultaneously using Ettan IPGphor manifold or 02 strips using ceramic strip holders in single time. Ettan IPGphor 3 Control Software allows real time monitoring of the all first dimensional run with similar set of run parameters. Focussed proteins are resolved on medium size slab gel using SE 600 vertical unit (Amersham biosciences).
    • Current Status of Instrument:
      Fully functional
    • Charges including GST
    • Name of Analysis Price per Sample (in ₹)) Remarks
      First dimension gel electrophoresis 10000  (Industry / Enterpreneur)
      5000    (Research/Academic/Farmer)
      		 Nil
      Second dimension gel electrophoresis 10000  (Industry / Enterpreneur)
      5000    (Research/Academic/Farmer)
      		 Nil
    • Contact details with e-mail & phone:
    • Praveen Maurye, ARS (Biotechnology)
      Scientist (SG)
      Fishery Resource & Environmental Management (FREM)
      ICAR- Central Inland Fisheries Research Institute
      Barrackpore, Kolkata-700120, W.B.
      Telefax: 033 -2545 2061(O); Mob. 9831943294
      E- mail: Praveen.maurye@icar.gov.in; maurye_p@yahoo.com
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