Detection of Plant Nematodes by Serological Analyses

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A quick and reliable routine test to identify nematodes is of great importance. Morphological species differentiation is arduous and not entirely reliable because of intra-and interspecific variations in shape and size. Differentiation based on biochemical or serological entities should offer better prospects. Immunoassays have great applications in agriculture for diagnosing crop diseases, pesticides, and other natural compounds. Since the generation of anti-nematode antibodies was first reported, several researchers have demonstrated promising results with the use of polyclonal and monoclonal antibodies (mAbs).

Lifeasible is committed to helping our customers achieve effective and successful research. We provide convenient and guaranteed detection of plant nematodes with serological analyses. In addition, we deliver reliable results and reports on time to our customers worldwide.

Detection of Plant Nematodes by Serological Analyses

Using more sophisticated immunochemical techniques, it may be possible to distinguish nematodes with conventional antisera.
Monoclonal antibodies are expected to be better differentiation reagents. They typically recognize a single epitope with a concomitant reducing cross-reactivity. In addition, rare antibody specificities, who's potential reactivities never become manifest in vivo, may arise in vitro by cell fusion with single members from the antigen-reactive B cell pool.
Lifeasible provides both polyclonal and monoclonal antibodies to detect plant nematodes, which is a quick and reliable approach based on biochemical and serological analyses.

Fig.1 SDS-PAGE of plant nematodes protein followed by immunoblotting. (Schots A, et al., 1989)

Workflows of Serological Analyses

Steps	Operation Methods
Antigen Preparation and Immunization of Animal	Antigenic proteins are isolated from plant nematodes and conjugated to Keyhole Limpet Hemocyanin. The protein conjugate is precipitated on aluminum hydroxide for immunization. The protein is spotted on nitrocellulose and dried for 16 h at room temperature. Nitrocellulose is mixed with PBS to pass it through an injection needle. The protein is subjected to preparative SDS-PAGE in cells.
Preparation of Monoclonal Antibodies	Cell fusions are carried out with cells; then, cell culture supernatants are screened for the presence of specific antibodies using an ELISA. Positive hybrid cell lines are cloned by limiting dilution. And heavy chain analysis is carried out with concentrated culture supernatants in a double diffusion assay.
Determination of Cross-Reactivity	Homogenates of other plant nematode species are obtained to determine cross-reactivity. Incubation with antibodies and visualization is performed.
Calculation of Binding Constants	Antibody binding constants are calculated from the results of a direct ELISA.
Electrophoresis and Blotting	Two-dimensional electrophoresis is carried out with a certain protein equivalent per gel. Immunoblotting of gels is performed by electro transfer of the proteins from the gel to an immobilon membrane.

Lifeasible has extensive experience and expertise in plant science. We are committed to providing you with timely and high-quality deliverables. At the same time, we guarantee the cost-effectiveness, completeness, and simplicity of the report. If you are interested in our services or have any questions, please feel free to contact us or make an online inquiry.

Reference

Schots A, et al. (1989). "Serological differentiation of the potato-cyst nematodes Globodera pallida and G. rostochiensis: II. Preparation and characterization of species-specific monoclonal antibodies." Hybridoma. 8 (4), 401-13.

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