Biotin (vitamin H) ELISA

Name: Biotin (vitamin H) ELISA
SKU: DEIA-Z77801
Rating: 5 (1 reviews)

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

COA

SDS

Datasheet

Summary

Sample

serum, plasma, urine and milk

Detection Method

cELISA

Intended Use

This assay is intended for the quantitative determination of biotin in serum, plasma, urine and milk.

Contents of Kit

1. PLATE: Microtiter plate, pre-coated, 12 × 8 wells
2. WASHBUF: Wash buffer concentrate, 10 × 2 × 100 ml
3. CONJ: Conjugate, ready-to-use 1 × 13 ml
4. STD: Standards, ready-to-use, 1 × 6 vials
5. CTRL A: Control, ready-to-use, 1 × 1 vial
6. CTRL B: Control, ready-to-use, 1 × 1 vial
7. SAMPLEBUF: Sample dilution buffer, ready-to-use, 1 × 30 ml
8. SUB: Substrate, ready-to-use, 1 × 15 ml
9. STOP: Stop solution, ready-to-use 1 × 15 ml
10. FOL: Lightproof foil to cover the microtiter plate, 3 × 1 piece

Storage

All other test reagents are ready-to-use. Test reagents are stable until the expiry date (see label) when stored at 2–8 °C.

Precision

Repeatability (Intra-Assay); n = 40
The repeatability was assessed with 2 serum samples under constant parameters (same operator, measurement system, day and kit lot).

Reproducibility (Inter-Assay); n = 15
The reproducibility was assessed with 2 serum samples under varying parameters (different operators, measurement systems, days and kit lots).

Detection Range

Samples with concentrations above the measurement range (> 1 100 ng/l) can be further diluted with sample dilution buffer and re-assayed. Please consider this greater dilution when calculating the results.
Samples with concentrations lower than the measurement range (= LoQ, 48,1 ng/l) cannot be clearly quantified.

Sensitivity

The following values have been estimated based on the concentrations of the standard without considering possibly used sample dilution factors.
Limit of blank, LoB 25 ng/l
Limit of detection, LoD 32.4 ng/l
Limit of quantitation, LoQ 48.1 ng/l
Measuring range: 48,1–1 100 ng/l

General Description

Biotin (vitamin H) is present in bacteria, funghi, plants and animals. In food, the major part of biotin is covalently bound to protein, leaving only a minor fraction freely available. During digestion, biocytin (biotinyl-lysine) is released from the proteins and can be resorbed as easily as biotin from the intestinal tract. Afterwards, biotin is released from biocytin by the enzyme biocytinase in erythrocytes and plasma. It is then available as prosthetic group for a series of biotin-dependent enzymes. The daily requirements of biotin are difficult to estimate because a healthy intestinal flora supplies a major part of biotin by endogenous synthesis. The generally recommended daily dose for adults is 100–200 μg. Chronic hemodialysis patients show clear improvements of neuropathological status and glucose metabolism when supplemented with biotin in a milligram range. Biotin deficiency can be caused by e. g. destruction of the intestinal flora or extreme diets (e. g. frequent consumption of raw eggs). It can lead to dermatitis, hair loss, anorexia, mucular hypotonia, depression and reproduction problems.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

The PCR-HCR dual signal amplification strategy for ultrasensitive detection of Escherichia coli O157:H7 in milk

LWT-FOOD SCIENCE AND TECHNOLOGY

Authors: Liang, Taobo; Wu, Xin; Chen, Bolu; Liu, Ju; Aguilar, Zoraida P.; Xu, Hengyi

Abstract

In this study, we established a novel polymerase chain reaction-hybridization chain reaction (PCR-HCR) dual signal amplification assay for sensitive detection of Escherichia coil O157:H7 in milk. In this assay, a blocker forward primer and a biotin reverse primer based on E. coil O157:H7 specific fliC(h7), gene were specifically designed for PCR amplification resulting in a single-double stranded DNA with biotin (ss-dsDNA similar to biotin). These PCR products, ss-dsDNA similar to biotin, were captured by magnetic beads (MBs) decorated with streptavidin (SA) forming the ss-dsDNA similar to biotin + SA-MBs. The ssDNA on the PCR products opened the hairpin probe (H1 and H2) to trigger the HCR for secondary signal amplification. This is the first to use HCR to directly amplify PCR products through a novel blocker primer. With the help of two nucleic acid dual signal amplification techniques, PCR-HCR analysis can effectively improve the detection sensitivity and the limit of detection is 7.2 x 10(1) CFU/mL. In addition, this assay has been successfully used to detection target bacteria in skim milk spiked samples.

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

Abstract

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.