Generation and Application of a Novel InsP₃R₁ Mono-Antibody from Mouse

<div><p>Inositol 1, 4, 5-Trisphosphate Receptor (InsP₃R) is an intracellular Ca²⁺ release channel, which widely participates in cellular processes. Three isoforms of InsP₃R were identified as InsP₃R₁, InsP₃R_2, and InsP₃R₃. They share 60–0% protein sequence homology and form a channel in a manner of homotetramer or heterotetramer. Several InsP₃R isoform-specific rabbit antibodies have been generated to distinguish their localization and functions. However, there is no report of such a valid antibody raised from other species. In his article, we prepare a mouse monoclonal antibody against a synthetic peptide with rat InsP₃R₁-specific carboxyl terminus sequence. This monoclonal antibody of InsP₃R₁ (R1-mAb) was purified and characterized as IgG2b, which can recognize InsP₃R₁ by Western-blot (WB) analysis and immunoprecipitate (IP) InsP₃R₁ from moue brain lysate tested. Applied in immunofluorescent (IF) and immunohistochemical (IHC) assays, this antibody and rabbit polyclonal antibody could give the consistent results in SH-SY5Y cells, human sperm, and mouse brain paraffin section. In summary, we generate a mouse InsP₃R₁-specific IgG 2b antibody identifying InsP₃R₁ in WB, IF, IHC, and IP analysis, which provides a possible choice for detection of InsP₃R₁, especially in application of co-localization analysis with other InsP₃R isoforms or other proteins. </p></div

Quantum-Dot-Based Lateral Flow Immunoassay for Detection of Neonicotinoid Residues in Tea Leaves

Neonicotinoid insecticides are commonly used for pest control on tea plantations as a result of their broad-spectrum activity. However, neonicotinoid residues released from tea leaves into tea infusions pose a dietary risk to consumers. Therefore, a rapid, sensitive, and reliable on-site detection method for neonicotinoids is needed. We developed a quantum-dot-based fluorescent lateral flow immunochromatographic strip (LFICS) combined with a broad-specific antibody for detection of typical neonicotinoids (imidacloprid, imidaclothiz, and clothianidin), with sensitivities [50% inhibitory concentration (IC₅₀)] of 0.104–0.33 ng/mL and visual detection limits of 0.5–1 ng/mL. The strip assay could be completed in less than 30 min. Using the LFICS to analyze spiked tea samples (green tea, black tea, and oolong tea), the average recovery of the three neonicotinoids ranged between 71 and 111%, with the coefficient of variation below 12%. The results from the LFICS tests for field samples were consistent with results from ultraperformance liquid chromatography–tandem mass spectrometry. The newly developed strip is a useful tool for the on-site detection of neonicotinoid residues in tea