In vitro diagnostics (IVD) is a promising and powerful tool used in the diagnosis of diseases, infections detecting, disease screening, therapeutic monitoring, as well as the recognition of allergies and measurement of hormones or biological markers in serum. Creative Biolabs has seasoned experienced in IVD productions and has launched a wide range of diagnosis kits, especially western blot ones, for medical studies and clinical trials.

Western Blot

Western blot (WB) is a commonly used technique in molecular and cell biology, as well as in protein engineering. Through a WB assay, researchers can separate and identify specific proteins from a complex mixture of protein extractions within cells. There are three main parts to accomplish this task involving separation by molecular size, transfer to a solid support through gel electrophoresis, and incubating target protein with appropriate labels of primary and secondary antibody to visualize.

(a) Assembled rack for gel solidification; (b) Add gel solution using a transfer pipette; (c) Add running buffer to the electrophorator; (d) Add samples and molecular marker to the gel, after removing the combs. Fig.1 (a) Assembled rack for gel solidification; (b) Add gel solution using a transfer pipette; (c) Add running buffer to the electrophorator; (d) Add samples and molecular marker to the gel, after removing the combs. (Mahmood., 2012)

WB is also referred to immunoblotting because this approach makes use of antibodies detecting particular proteins from the mixture. In this assay, the labeled antibody against specific protein is performed to identify the desired protein, so it is a test with specificity. In a regular WB process, the followed protocols are included:

  • Cell lysis to extract protein samples
  • Gel preparation: SDS-PAGE
  • Electrophoresis and electrotransfer
  • BSA blocking
  • Antibody incubations (Treatment with primary antibody and then secondary antibody)
  • Imaging and analysis

Western blot profiles obtained with sera from patients infected by various species of Trichinella.Fig.2 Western blot binding procedure.

WB-based Kits Development

Many techniques of antibody assays, such as indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA), have been developed is reviewed in the past. Though sensitive, these methods are also subject to cross-reactions that make weakly positive results difficult to be interpreted.

Western Blot Based Kits DevelopmentFig.3 Western blot profiles obtained with sera from patients infected by various species of Trichinella. (Year., 2003)

In Creative Biolabs, we have developed fast western blot-based kits for IVD to accelerate the experimental process. The optimized reagents and simplified protocols enable a more efficient, accuracy, and reliability outcome. Furthermore, our IVD products are equipped with other extraordinary advantages. They are convenient, simple and easy to operate, no extra expensive consumables and hardware required. The kits are stable in proper storage and contain all elements needed in WB to block, probe and develop a blot.

References

  1. Mahmood, T., (2012). “Western blot: technique, theory, and trouble shooting.” N Am J Med Sci, 4(9), 429-434.
  2. Year, H., (2003). “Development and evaluation of a Western blot kit for diagnosis of human trichinellosis.” Clin Diagn Lab Immunol, 10(5), 793-796


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