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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Determination of the Relative Cell Surface and Total Expression of Recombinant Ion Channels Using Flow Cytometry
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Determination of the Relative Cell Surface and Total Expression of Recombinant Ion Channels Using Flow Cytometry

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Microsphere surface protein determination using flow cytometry.

Travis A Woods1, Steven W Graves, John P Nolan

  • 1Los Alamos National Laboratory, Los Alamos, New Mexico, USA.

Current Protocols in Cytometry
|September 5, 2008
PubMed
Summary
This summary is machine-generated.

This study introduces a new method to quantify surface proteins on microspheres using the CBQCA dye. This technique enables accurate measurement of proteins, even without known binding partners, via flow cytometry.

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Area of Science:

  • Bioconjugation and Surface Chemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Accurate quantification of surface-bound proteins is crucial for various applications, including diagnostics and drug delivery.
  • Existing methods often require known binding partners or specific surface chemistries, limiting their applicability.

Purpose of the Study:

  • To describe an extrinsic staining protocol for quantifying surface proteins on microspheres.
  • To introduce the use of the amine-reactive CBQCA dye for this purpose.
  • To enable quantification of proteins lacking known binding partners.

Main Methods:

  • Utilizing the amine-reactive CBQCA dye for extrinsic staining of microspheres.
  • Employing flow cytometry for accurate protein quantification.
  • Developing a protocol applicable to proteins without defined binding partners.

Main Results:

  • Successfully established an extrinsic staining protocol using CBQCA dye.
  • Demonstrated accurate quantification of surface proteins on microspheres.
  • Validated the method for proteins lacking known binding partners.

Conclusions:

  • The CBQCA dye offers a versatile tool for quantifying surface proteins on microspheres.
  • This protocol enhances the ability to analyze protein-coated microspheres in diverse research areas.
  • Flow cytometry combined with CBQCA staining provides a robust method for protein characterization.