EdU Imaging Kits (HF488)
![mRNA synthesis](/media/diy/images/page/figure1-mrna.png)
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
![Tyramine Signal Amplification (TSA)](/media/diy/images/page/figure2-01.png)
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
![screening library](/media/diy/images/page/figure3-01.png)
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
![Cell Counting Kit-8 (CCK-8)](/media/diy/images/page/CCK-8.jpg)
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
![SYBR Safe DNA Gel Stain](/media/diy/images/page/SYBR Safe DNA Gel Stain.png)
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
![Inhibitor Cocktails](/media/diy/images/page/Inhibitor Cocktails.jpg)
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
测量细胞增殖和细胞周期是评估细胞健康、确定遗传毒性和评估药物药效的基本方法,常用的方法是直接测量DNA 的合成。在以往的实验中常用方法包括掺入放射性核苷(3H-thymidine)或BrdU等。而EdU Imaging Kits (HF488)则采用了一种新的方法:点击化学-CuAAC(铜催化的叠氮化物-炔烃环加成反应),使用该反应可直接测量在细胞周期S期的DNA合成。
在基于BrdU的检测中,需要额外的DNA变性(通常使用HCl、加热或用DNase消化)以使BrdU暴露于抗BrdU抗体。胸腺嘧啶核苷类似物EdU(5-ethynyl-2’-deoxyuridine)是BrdU的理想替代品,因为它不受苛刻的DNA变性条件的影响,EdU可以在DNA合成过程中掺入DNA链中。EdU的炔基是一种生物惰性基团,可以通过CuAAC反应与染料的叠氮基发生极强的选择性反应,从而生成1, 2, 3-三唑产物。EdU和HyperFluor™ 488 azide具有生物学上独特的基团,其连接后可以荧光标记增殖细胞的DNA,具有低背景和高检测灵敏度。这种CuAAC反应可在极温和的条件下提供出色的区域选择性和定量转化。该反应是高效的,并且可以通过流式细胞术或荧光显微镜定量地使用。
EdU Imaging Kits (HF488) 采用的是HyperFluor™ 488 azide与EdU进行连接后荧光标记增殖细胞的DNA,可通过荧光显微镜进行观察(HyperFluor™ 488 azide最大激发波长为496 nm,最大发射波长为516 nm)。
Components | 50-500 Tests |
EdU (Component A) | 5 mg |
HyperFluor™ 488 azide (Component B) | 25 μL |
DMSO (Component C) | 4 mL |
10X EdU Reaction Buffer (Component D) | 4 mL |
CuSO4 (100 mM Aqueous Solution) (Component E) | 1 vial |
EdU Buffer Additive (Component F) | 400 mg |
Hoechst 33342 (10 mg/mL in Water) (Component G) | 35 μL |
Store the kit at -20ºC away from light and moisture, stable for 1 year. |