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Related Experiment Videos

Tissue factor mRNA quantitation without prior monocyte isolation.

S Javorschi1, S Labrouche, G Freyburger

  • 1Laboratoire Hématologie, Hôpital Pellegrin, Bordeaux, France. sandrine.javorschi@hematologie.u-bordeaux2.fr

Biotechniques
|June 27, 2000
PubMed
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Quantifying monocyte tissue factor (TF) mRNA using CD11b as a reference gene provides a reliable method for assessing monocyte activation in various diseases. This approach overcomes limitations of previous techniques for evaluating coagulation processes.

Area of Science:

  • Biochemistry
  • Immunology
  • Molecular Biology

Background:

  • Monocyte tissue factor (TF) quantitation is crucial for understanding coagulation in diseases.
  • Current methods for assessing TF expression in monocytes face technical challenges, hindering reliable evaluation.
  • Early detection of monocyte activation is vital for disease management.

Purpose of the Study:

  • To develop a standardized and accurate method for quantifying monocyte TF expression.
  • To identify a suitable reference gene for normalizing TF mRNA levels in monocytes.
  • To establish a sensitive assay for early detection of monocyte activation.

Main Methods:

  • Quantitative reverse transcription polymerase chain reaction (RT-PCR) for TF mRNA.
  • Automated electrophoresis for sensitive product detection.

Related Experiment Videos

  • Validation of CD11b as a reference gene for monocyte TF mRNA standardization.

    • Main Results:

    • CD11b gene expression was found to be suitable for standardizing TF mRNA quantitation.
    • CD11b exhibits stable expression during inflammation and similar levels to target mRNA.
    • The proposed method allows TF expression evaluation without further monocyte purification.

    Conclusions:

    • CD11b is a validated reference gene for accurate monocyte TF mRNA quantitation.
    • This method offers a rapid, sensitive, specific, and accurate approach to evaluate monocyte TF expression.
    • The findings facilitate reliable assessment of coagulation involvement in diseases.