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Related Experiment Videos

Purification and characterization of rabbit tissue factor.

L V Rao1, A D Hoang

  • 1Department of Medicine, University of California, San Diego, La Jolla 92093.

Thrombosis Research
|October 1, 1989
PubMed
Summary

Researchers developed a rapid, one-step method to purify rabbit brain tissue factor using an anti-human tissue factor IgG immunoadsorbent column. This technique yields a highly pure antigen for antiserum production and molecular characterization.

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Area of Science:

  • Biochemistry
  • Immunology
  • Protein Purification

Background:

  • Tissue factor (TF) is a critical initiator of the coagulation cascade.
  • Rabbit TF is a valuable model for studying TF function and regulation.
  • Existing purification methods for TF can be laborious and yield low purity.

Purpose of the Study:

  • To develop an efficient and rapid method for purifying rabbit brain tissue factor.
  • To generate a high-titer, monospecific polyclonal antiserum against rabbit TF.
  • To characterize the molecular properties of purified rabbit TF.

Main Methods:

  • Partial purification of rabbit TF using an anti-human TF IgG affinity column.
  • Further purification of rabbit TF via SDS-polyacrylamide gel electrophoresis (SDS-PAGE).

Related Experiment Videos

  • Generation of polyclonal antiserum using purified rabbit TF as antigen.
  • Immunostaining of rabbit brain extracts and purified TF.
  • Coupling of anti-rabbit TF IgG to Affi-Gel-15 for immunoadsorption.

    • Main Results:

    • Essentially homogeneous rabbit TF was obtained after sequential purification.
    • A high-titer, monospecific polyclonal antiserum against rabbit TF was successfully raised.
    • Immunostaining revealed a single major band at 45 kD, corresponding to rabbit TF apoprotein.
    • The anti-rabbit TF IgG immunoadsorbent column provided a rapid, one-step purification of large amounts of rabbit brain TF.

    Conclusions:

    • A novel and efficient immunoadsorbent strategy was established for rabbit TF purification.
    • The developed antiserum is suitable for identifying and quantifying rabbit TF.
    • This method significantly simplifies and accelerates the purification of rabbit TF for research purposes.