Biotin ELISA Kit (DEIA281)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
food
Species Reactivity
N/A
Intended Use
Enzyme Immunoassay for the Quantitative Determination of Biotin (Vitamin H) in Food
Contents of Kit
1. Microtiter plate
2. Biotin Standards(0; 1; 2.5; 5; 10; 25 ng/mL)
3. Conjugate(Biotin-Alkaline Phophatase)
4. Substrate Solution (PNPP)
5. Stop Solution (1M NaOH)
6. Standard/Sample Diluent (PBS)
7. Washing Solution(PBS + Tween 20)
8. plastic foils
Storage
Stored at 2-8°C. Expiry data are found on the labels of the bottles and the outer package. For more detailed information, please download the following document on our website.
Precision
The Intra-assay variation of the Biotin test was determined to 3%.
Sensitivity
0.5 ng/mL

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References


Pine wood nematode (PWN)-secretory antigen 571 as a biomarker for the PWN and its monoclonal antibodies for detecting PWN and its infected pine tree

JOURNAL OF ASIA-PACIFIC ENTOMOLOGY

Authors: Osabutey, Angelina F.; Kim, A-Young; Nguyen, Phoung; Ha Oh, Kyu; Han, Hyerim; Koh, Young Ho

The biochemical characteristics of pine wood nematode (PWN)-secretory antigen 571 (PWN-SA571) identified from the PWN secretome and PWN-infected pine tree extracts (PWNIPT) were investigated. PWN-SA571 has homologs in various nematodes in diverse families and a signal peptide at the amino-terminal end. Since it was not possible to express the full-length PWN-SA571 protein, two peptides showing high antigenicity were synthesized and used as antigens for generating monoclonal antibodies (Mabs). Mab-secreting fusion hybridoma cell lines were selected based on immunoreactivity to free peptide and BSA-conjugated peptide (BSA pep) antigens by enzyme-linked immunosorbent assay for establishing Mab-secreting hybridoma lines. Ascites containing PWN-SA571-specific Mabs showed stronger immunoreactivity to PWNIPT than to healthy pine tree extracts. In addition, PWN-SA571-specific Mabs could recognize less than 20 ng of BSA pep in lateral flow assays. Furthermore, purified biotin-conjugated PWN-SA571-specific Mab IgG or IgM were able to detect BSA pep (< 15 ng) and PWN extracts (< 600 ng). Taken together, these results demonstrate that PWN-SA571-specific Mabs could detect PWN or PWNIPT extracts by ELISA, LFA, or dot blot analysis.

lincRNA_Tc13743.2-miR-133-5p-TcGSTm02 regulation pathway mediates cyflumetofen resistance in Tetranychus cinnabarinus

INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY

Authors: Feng, Kaiyang; Liu, Jie; Wei, Peng; Ou, Shiyuan; Wen, Xiang; Shen, Guangmao; Xu, Zhifeng; Xu, Qiang; He, Lin

Differential expression of metabolic detoxification enzymes is an important mechanism involved in pesticide/ acaricide resistance of mite pests. The competing endogenous RNA hypothesis offers a new opportunity to investigate post-transcriptional regulation of those genes. In this study, 4454 long non-coding RNAs were identified in the carmine spider mite Tetranychus cinnabarinus by transcriptome sequencing. Software-based predictions indicated that a long intergenic non-coding RNA, (lincRNA)_Tc13743.2 and a detoxification enzyme gene, TcGSTm02, both contained a microRNA (miR-133-5p) response element. Over-expression of lincRNA_Tc13743.2 and TcGSTm02 were detected in a cyflumetofen-resistant T. cinnabarinus strain (CyR), whereas down-regulation of miR-133-5p was observed in this strain. Conversely, up-regulation of miR-133-5p could inhibit TcGSTm02 expression levels, and both lincRNA_Tc13743.2 and TcGSTm02 were significantly enriched in miR-133-5p biotin-avidin pull-down assays. RNA-binding protein immunoprecipitation assay showed that lincRNA_Tc13743.2 and TcGSTm02 bound to a silencing complex containing miR-133-5p. Moreover, a luciferase reporter assay based on a human cell line revealed that over-expression of lincRNA_Tc13743.2 could significantly reduce the inhibition exerted by miR-133-5p through the TcGSTm02 3 ' UTR. In addition, co -localization of lincRNA_Tc13743.2 and miR-133-5p was detected using fluorescence in situ hybridization, suggesting that lincRNA_Tc13743.2 interacts directly with miR-133-5p in spider mites. More importantly, silencing the expression of lincRNA_Tc13743.2 significantly reduced the expression levels of TcGSTm02 and increased the sensitivity of spider mites to cyflumetofen. Our data show that lincRNA_Tc13743.2 up-regulates TcGSTm02 expression by competing for miR-133-5p binding, demonstrating that a lincRNA_Tc13743.2-miR-1335p-TcGSTm02 pathway mediates cyflumetofen resistance in mites.

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