Cell Proliferation EdU Image Kit (Green Fluorescence) is a novel alternative to the BrdU assay. EdU (5-ethynyl-2´-deoxyuridine) is a nucleoside analog of thymidine and is incorporated into DNA during active DNA synthesis. Comparing to BrdU assays, the EdU-Click Assays are not antibody based and therefore do not require DNA denaturation (typically using HCl or heat or digestion with DNase) for detection of the incorporated nucleoside. Detection is based on a click reaction, a copper-catalyzed covalent reaction between an azide and an alkyne, is complete within 30 minutes.
• Optimized negative control is provided to rule out false positives
•No antibody needed.
• No denaturation steps, preservation of cell morphology and DNA integrity.
• Simple, reliable and time-saving than traditional methods.
• Proprietary AbFluor 488 azide (Ex/Em = 501/525 nm)-good photostability and minimizing fluorescence quenching.
• Optimized for fluorescent microscopy.
Storage instructions
Stable for at least 12 months at recommended temperature from date of shipment. Gel pack with blue ice.
Shipping
Gel pack with blue ice.
Precautions
The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.
Additional Information
Background
The detection of cell proliferation is of utmost importance for assessing cell health, determining genotoxicity or evaluating anticancer drugs. Until now, measuring DNA synthesis directly is most accurate method of doing it, normally performed by incorporation of the nucleoside analog like [3H] thymidine or 5-bromo-2’-deoxyuridine to cells during replication, and then detected or visualized by autoradiography or with an anti-BrdU-antibody respectively.
Prostate cancer is the most common tumor in men worldwide, seriously threatening the health of older men, and 5-methylcytosine (m5C) RNA modification has been shown to have a significant impact on the development and progression of various tumors. However, as the most critical methyltransferase for m5c RNA modification, the role of the NSUN members (NSUN1-7) in prostate cancer is unclear.
MiRNAs have been found to be involved in the regulation of ovarian function as important post-transcriptional regulators, including regulators of follicular development, steroidogenesis, cell atresia, and even the development of ovarian cancer. In this study, we evaluated the regulatory role of gga-miR-449b-5p in follicular growth and steroid synthesis in ovarian granulosa cells (GCs) of laying hens through qRT-PCR, ELISAs, western blotting and dual-luciferase reporter assays, which have been described in our previous study. We demonstrated that gga-miR-449b-5p was widely expressed in granulosa and theca layers of the different-sized follicles, especially in the granulosa layer. The gga-miR-449b-5p had no significant effect on the proliferation of GCs, but could significantly regulate the expression of key steroidogenesis-related genes (StAR and CYP19A1) (p < 0.01) and the secretion of P4 and E2 (p < 0.01 and p < 0.05). Further research showed that gga-miR-449b-5p could target IGF2BP3 and downregulate the mRNA and protein expression of IGF2BP3 (p < 0.05). Therefore, this study suggests that gga-miR-449b-5p is a potent regulator of the synthesis of steroid hormones in GCs by targeting the expression of IGF2BP3 and may contribute to a better understanding of the role of functional miRNAs in laying hen ovarian development.