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DAPI染色液(1mg/ml)

一键复制产品信息

货号:AWC0292

价格: ¥190

规格: 1ml 10ml

  • 产品概述
  • DAPI染色液(1mg/ml)

    产品简介:

    DAPI染色液(DAPI Staining Solution)是适用于常见细胞和组织细胞核染色的染色液。DAPI,即2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride,也称DAPI dihydrochloride,分子式为C16H15N5·2HCl,分子量为350.25,是可以穿透细胞膜的蓝色荧光染料,和双链DNA结合后可以产生比DAPI自身强20多倍的荧光,灵敏度高于EB。

    DAPI染色常用于细胞凋亡检测,染色后用荧光显微镜观察或流式细胞仪检测。DAPI也常用于普通的细胞核染色以及某些特定情况下的双链DNA染色。DAPI的最大激发波长为340nm,最大发射波长为488nm。DAPI和双链DNA结合后,最大激发波长为364nm,最大发射波长为454nm。DAPI染色液是浓缩的储存液,稀释后使用,一般推荐工作浓度为0.5~10μg/ml,用于固定细胞或组织的细胞核染色。

    自备材料:

    1、 荧光显微镜

    2、 蒸馏水

    3、 微量移液器

    4、 PBS或生理盐水

    操作步骤(仅供参考):

    1、 根据实验具体要求,用无菌去离子水稀释到自己所需浓度,即为DAPI染色工作液。细胞染色时,一般推荐工作浓度为0.5~10μg/ml。

    2、 对于细胞或组织样品,固定后冲洗去除固定剂。如果需要进行免疫荧光染色,则先进行免疫荧光染色,染色完毕后再按后续步骤进行DAPI染,如果不需要进行其它染色,则直接进行后续的DAPI染色。对于贴壁细胞或组织切片,加入少量DAPI染色液,覆盖住样品即可。对于悬浮细胞,至少加入待染色样品3倍体积以上的DAPI染色液,充分混匀。

    3、 室温放置5~8min。

    4、 轻轻吸除DAPI染色液。

    5、 用无菌的PBS或生理盐水清洗2~3次,每次3~5min。

    6、 直接在荧光显微镜下观察或封片后荧光显微镜下观察。

    染色结果:细胞发生凋亡时,会看到凋亡细胞的细胞核呈致密浓染,或呈碎块状致密浓染。          

    注意事项:

    1、 DAPI染色液(1mg/ml)应稀释至合适的浓度后使用。

    2、 荧光染料都存在淬灭的问题,建议染色后尽快检测。

    3、 为减缓荧光淬片,可以使用抗荧光淬灭封片液。

    4、 避免反复冻融,否则容易失效。

    5、 DAPI对人体有一定刺激性,请注意适当防护。

    6、 为了您的安全和健康,请穿好实验服并佩戴一次性手套和口罩操作。

    7、 本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。

    产品组成
    名称货号规格storage
    DAPI染色液(1mg/ml)AWC0292-1ml1ml-20℃,避光
    DAPI染色液(1mg/ml)AWC0292-10ml10ml-20℃,避光

    注意:

    1.本产品仅供科研使用。请勿用于医药、临床诊断或治疗。食品及化妆品等用途。请勿存放于普通住宅区。

    2.为了您的安全和健康,请穿好实验服并佩戴一次性手套和口罩操作。

    3.实验结果可由多种因素影响,相关处理只限于产品本身,不涉及其他赔偿。


    参考文献 (20)

    ADVANCED FUNCTIONAL MATERIALS IF:19

    Bioengineered tracheas have shown considerable potential in tracheal injury repair; however, their practical value is limited by challenges in tracheal cartilage regeneration, and postoperative tracheal stenosis remains a common issue. Here, inspired by the 3-layer structure of the trachea and the multi-segmental characteristics of its cartilage, a multilayered bioengineered tracheal scaffold (named Sd@d-ECM/BMSCs/SilMA) with a microgroove structure is designed in this study to repair tracheal defects. In this design, the microgrooved surface of the methacrylated silk fibroin (SilMA) hydrogel provides spatial guidance for the directional growth of bone marrow mesenchymal stem cells (BMSCs) and enhances their adhesion and proliferation. The extracellular matrix of the decellularized cartilage scaffold offers the necessary microenvironment and mechanical support for BMSCs to differentiate into cartilage. Under the influence of the dual-layer structure (inner and outer), the middle-layer BMSCs can undergo stable chondrogenic differentiation without any inducing agents. Sd@d-ECM/BMSCs/SilMA effectively promotes tracheal cartilage formation in a rabbit defect model, reduces the incidence of tracheal stenosis, and substantially improves respiratory function. Sd@d-ECM/BMSCs/SilMA not only confirms the successful construction of microgroove structures on the surface of the SilMA hydrogel and the effective loading of BMSCs but also demonstrates significant experimental value in tracheal cartilage repair and regenerative medicine.

    Biomaterials Research IF:9.6

    Although tea consumption has been suggested to affect kidney stone formation, epidemiological evidence remains inconsistent, and the underlying molecular mechanisms are unclear. To assess the association between tea intake and kidney stone risk, we initially conducted a prospective cohort analysis of 481,393 participants from the UK Biobank and a 2-sample Mendelian randomization (MR) analysis. Our findings revealed that heavy tea drinkers (>5 cups/day) had a significantly reduced risk of kidney stones (hazard ratio: 0.79, 95% confidence interval [CI]: 0.72 to 0.86, P < 0.001), and MR analyses confirmed a causal association (inverse variance weighted OR: 0.45, 95% CI: 0.32 to 0.62, P < 0.001). We next explored the effect of epigallocatechin gallate (EGCG), the main bioactive component in tea, on calcium oxalate (CaOx) stone formation. EGCG was found to inhibit the glucose-regulated protein 94/phosphatidylinositol 3-kinase/protein kinase B (GRP94/PI3K/AKT) pathway in human proximal renal tubular epithelial cells, thereby attenuating CaOx crystal-induced oxidative stress and inflammation, and inhibiting crystal-cell adhesion. This finding aligned with the observation that the activated GRP94/PI3K/AKT pathway was positively associated with inflammation-related molecules in renal papillary tissues of CaOx stone formers. Moreover, to enhance renal targeting and therapeutic potential, we synthesized cell membrane-coated EGCG-loaded poly(lactic-co-glycolic acid) (TP-EGCG) nanoparticles, which enhanced renal EGCG delivery and substantially reduced CaOx crystal deposition in a mouse model of CaOx nephrolithiasis. In conclusion, tea consumption protects against kidney stone formation, an effect exerted by EGCG through the GRP94/PI3K/AKT axis, and our novel TP-EGCG nanoparticles show strong potential for targeted prevention and treatment.

    INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES IF:8.5

    Chemodynamic therapy (CDT) is a highly promising cancer treatment strategy. However, its clinical application is severely limited due to insufficient H 2 O 2 levels. To overcome this critical issues, based on self-supply H 2 O 2 strategy this study designed a multifunctional nano-reactor hyaluronic acid (HA)-cinnamaldehyde( CA )Schiff base(HA-CA)@hollow CaO 2 (HMCaO 2 )/Copper doped luteolin carbon dots (CuLCDs) (abbreviation:HACOCLC). HACOCLC based on self-supplied H 2 O 2 /O 2 strategy to enhance the anti-tumor effect. HACOCLC preferentially enrich at the tumor site under the tumor targeting effect of HA. pH stimulation triggered the release of HMCaO 2 and CuLCDs from HACOCLC. HMCaO 2 hydrolyzed to produce H 2 O 2 , Ca 2+ and O 2 , H 2 O 2 compensated substrates for CDT, O 2 down-regulates hypoxia-inducing factor (HIF-1α) and programmed death ligand 1(PD-L1) to consolidate the therapeutic effect, and Ca 2+ entered mitochondria to cause calcium overload and induced cell death. CuLCDs underwent Fenton-like reaction to produce •OH (CDT effect) and O 2 , which further increased the O 2 level. CuLCDs has near-infrared light (808 nm) thermal conversion (conversion efficiency was 40.3 %) for photothermal therapy (PTT), Cu 2+ depletion of glutathione amplified the level of reactive oxygen species (ROS) and improved the therapeutic effect. HACOCLC self-supplied H 2 O 2 and O 2 to enhance CDT, and effectively inhibited tumor growth through the combined action of CDT/PTT/ calcium overload.

    PHYTOTHERAPY RESEARCH IF:6.3

    Traumatic brain injury (TBI) is a severe condition with a high mortality rate, affecting multiple organs, including the gastrointestinal (GI) tract. Ghrelin is a brain-gut peptide that regulates the microbiota-brain-gut axis, facilitating communication between the GI tract and the central nervous system. This study aimed to investigate the role of ferulic acid (FA) in regulating Ghrelin to improve TBI and GI disorders (GID) induced by controlled cortical impact (CCI). This study used CCI as the in vivo TBI model and scratch-induced injury of primary astrocytes as the in vitro TBI model. The role and mechanism of FA modulation of Ghrelin in ameliorating TBI and GID were explored using multi-omics and network pharmacology analyses. In vivo, results revealed that FA is the main active component of the Guanxin II compound and mimics its function. Significant improvement in GI hypomotility and brain injury was observed in the FA group compared to the CCI group. Concurrently, FA ameliorated intestinal barrier impairment triggered by CCI-induced reduction in the expression of Ghrelin and reduces the inflammatory response. Furthermore, 16S rRNA results indicated that CCI-induced TBI worsened gut microflora imbalance via the brain-gut axis, while gut dysbiosis aggravated brain injury. FA improved the dysbiosis of Bacteroidetes and Odoribacter mainly by targeting the Ghrelin-mediated inflammatory response. RNA-seq and network pharmacology analyses revealed that FA mainly affects inflammation-mediated pyroptosis pathways in the brain-gut axis. Additionally, experimental evidence demonstrated that FA reversed CCI-induced pyroptosis in rats and scratch injury-induced pyroptosis in astrocytes by promoting the binding of Ghrelin to GHSR, which suppressed the TLR4/NF-κB/NLRP3 pathway. Conclusively, FA could alleviate TBI and GID by promoting Ghrelin to regulate the microbiota-brain-gut axis inflammation via the Ghrelin/TLR4/NLRP3 pathway.

    Biology Direct IF:5.7

    Background Ischemia/reperfusion (I/R) injury is a severe brain disorder with currently limited effective treatments. This study aims to explore the role of N6-methyladenosine (m6A) modification and associated regulatory factors in I/R to identify potential therapeutic targets. Methods We utilized a middle cerebral artery occlusion (MCAO) rat model and SH-SY5Y cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to assess m6A levels and investigate the impact of METTL3 overexpression on long non-coding RNA (lncRNA) CRNDE expression. The effects of silencing lncRNA CRNDE on the interaction between YTHDC1 and ATG10 mRNA, as well as the stability of ATG10 mRNA, were evaluated. Additionally, apoptosis rates, pro-inflammatory and anti-inflammatory factor levels, ATG10 expression, and autophagic activity were analyzed to determine the effects of METTL3 . The reverse effects of YTHDC1 overexpression were also examined. Results MCAO rats and OGD/R-treated SH-SY5Y cells exhibited reduced m6A levels. METTL3 overexpression significantly inhibited lncRNA CRNDE expression. Silencing lncRNA CRNDE mitigated OGD/R-induced apoptosis and inflammation in SH-SY5Y cells, while enhancing autophagy and stabilizing ATG10 mRNA. METTL3 overexpression decreased cell apoptosis, reduced the levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and increased IL-10 secretion. Furthermore, METTL3 overexpression upregulated ATG10 expression and promoted autophagy. Conversely, lncRNA CRNDE overexpression negated these effects. Conclusion The inhibition of lncRNA CRNDE affects the interaction between YTHDC1 and ATG10 mRNA and stabilizes ATG10 mRNA, mediated by METTL3 overexpression. These findings suggest that targeting lncRNA CRNDE to reduce apoptosis, inhibit inflammation, increase ATG10 expression, and enhance autophagy could offer new therapeutic strategies for I/R injury.

    ACS Applied Nano Materials IF:5.3

    Carbon dots as drug carriers have received increasing attention in nanotherapy. However, the use of chemical small molecules or polymers as precursors to carbon dots often leads to toxicity, thus limiting their practical application. In contrast, herbs are biocompatible plants with complex active ingredients, making them attractive candidates for carbon dot precursors. In this paper, we prepared antitumor carbon dots (LuCDs) using a hydrothermal method and luteolin extract as a precursor. Furthermore, by modifying copper ions on the surface of LuCDs, we obtained Cu-LuCDs. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) analysis results showed that both LuCDs and Cu-LuCDs effectively inhibited the proliferation of CT26 colon cancer cells. Additionally, these modified carbon dots exhibited photothermal conversion properties not present in the precursor alone. Under 808 nm optical excitation, they achieved photothermal conversion powers of 55 and 56%, respectively. The Cu-LuCDs effectively decompose hydrogen peroxide, generating cytotoxic •OH and oxygen through Fenton-like reactions, thereby alleviating tumor hypoxia and inhibiting the expression of hypoxia-inducing factor HIF-1α. Immunofluorescence analysis confirmed that both LuCDs and Cu-LuCDs induced immune cell death and activated systemic immune activity. When combined with the immune adjuvant αPD-L1, Cu-LuCDs successfully ablated primary tumors and significantly inhibited the growth of metastatic tumors (distal tumors). The design and analysis of the antitumor activity of Cu-LuCDs provide important inspiration for the application of herbal carbon dots.

    Communications Biology IF:5.1

    Endometrial fibrosis is the main feature of intrauterine adhesion (IUA). The m6A methylation is involved in the process of fibrosis. However, the regulatory pathways involved in m6A methylation in endometrial fibrosis remain unclear. ALKBH5 is differentially low expressed in the endometrial tissues of IUA. Overexpression of ALKBH5 inhibits TGF-β1-induced fibrosis. In the in vivo experiment, ALKBH5 overexpression reduces the degree of endometrial fibrosis in rats. ALKBH5 regulates the m6A methylation level of FABP4 mRNA. FABP4 expression is inhibited by WT- ALKBH5 , but not by catalytically inactive MUT- ALKBH5 (H204A). The m6A reader IGF2BP2 targets FABP4 and affects FABP4 mRNA stability. Inhibition of FABP4 decreases the expression of fibrosis-related markers (α-SMA, collagen I, collagen III, and Fibronectin). In addition, serum lipid metabolism is disordered in IUA rats, and ALKBH5 overexpression could partially reverse the levels of differential lipid metabolites. In conclusion, ALKBH5 is differentially low expressed in IUA. ALKBH5 signaling regulates endometrial fibrosis through FABP4 mRNA m6A methylation and lipid metabolism. This finding can provide theory support for the potential treatment strategy development of IUA.

    LABORATORY INVESTIGATION IF:5

    Bladder cancer is a malignant tumor of the urinary system and is one of the most common cancers worldwide. Lipoxygenases are closely related to the development of various cancers. However, the relationship between lipoxygenases and p53/SLC7A11–dependent ferroptosis in bladder cancer has not been reported. Here, we aimed to investigate the roles and internal mechanisms of lipid peroxidation and p53/SLC7A11–dependent ferroptosis in the development and progression of bladder cancer. First, ultraperformance liquid chromatography-tandem mass spectrometry was performed to measure the metabolite production of lipid oxidation in patients’ plasma. The metabolic changes in patients with bladder cancer were discovered, revealing that stevenin, melanin, and octyl butyrate were upregulated. Then, the expressions of lipoxygenase family members were measured to screen out candidates with significant changes in bladder cancer tissues. Among various lipoxygenases, ALOX15B was significantly downregulated in bladder cancer tissues. Moreover, p53 and 4-hydroxynonenal (4-HNE) levels were decreased in bladder cancer tissues. Next, sh-ALOX15B, oe-ALOX15B, or oe-SLC7A11 plasmids were constructed and transfected into bladder cancer cells. Then, the p53 agonist Nutlin-3a, tert-butyl hydroperoxide, iron chelator deferoxamine, and the selective ferroptosis inhibitor ferr1 were added. The effects of ALOX15B and p53/SLC7A11 on bladder cancer cells were evaluated by in vitro and in vivo experiments. We revealed that knockdown of ALOX15B promoted bladder cancer cell growth, which was also found to protect bladder cancer cells from p53-induced ferroptosis. Furthermore, p53 activated ALOX15B lipoxygenase activity by suppressing SLC7A11. Taken together, p53 activated the lipoxygenase activity of ALOX15B via inhibiting SLC7A11 to induce ferroptosis in bladder cancer cells, which provided insight into the molecular mechanism of the occurrence and development of bladder cancer.

    INTERNATIONAL IMMUNOPHARMACOLOGY IF:4.8

    Knee osteoarthritis (KOA) is a chronic disease characterized by joint wear and cartilage degeneration. Current clinical treatments are based on symptomatic relief and are not effective in regenerating cartilage, and inflammation-induced cartilage damage accelerates the progression of osteoarthritis, making the protection of articular cartilage important for controlling the development of knee osteoarthritis. In this study, a biodegradable hydrogel (HA-Ca-Alg@Ica) loaded with Icariin (Ica) was prepared by in situ cross-linking of hyaluronic acid-calcium complex (HA-Ca) and sodium alginate (Alg-Na) for local sustained delivery of Ica. The hydrogel promoted chondrocyte proliferation and inhibited the degradation of cartilage matrix by regulating key factors (Wnt3a, β-catenin and GSK-3β) in the Wnt/β-catenin signaling pathway. In addition, the hydrogel reduced the expression of inflammatory factors, including IL-1β, IL-6, TNF-α, COX-2, and MMP13, leading to a reduction in inflammation and pain relief. In summary, this hydrogel containing Icariin has shown significant effects in reducing chondrocyte degradation and promoting chondrocyte proliferation, which can play a role in delaying osteoarthritis by protecting chondrocytes. These findings offer innovative prospects for the therapeutic management of knee osteoarthritis.

    Biomolecules IF:4.8

    Cerebral ischemia/reperfusion (I/R) injury is a devastating neurological disorder with limited treatment options. Emerging evidence suggests that the N6-methyladenosine (m6A) modification and its regulatory factors play pivotal roles in the pathophysiology of I/R. This study aimed to elucidate the function of METTL3-mediated m6A modification of the long non-coding RNA (lncRNA) AU020206 in ferroptosis during cerebral I/R injury and to identify potential molecular targets for neuroprotection. A murine model of middle cerebral artery occlusion/reperfusion (MCAO/R) and N2a cells subjected to oxygen–glucose deprivation/reoxygenation (OGD/R) were established to assess m6A levels and ferroptosis-related changes. Effects of METTL3 overexpression and lncRNA-AU020206 silencing on neuronal apoptosis, inflammation, and ferroptosis were investigated in vitro and in vivo. The interaction between lncRNA-AU020206 and YTHDC2 and the resulting regulation of SLC7A11 mRNA stability and GPX4 expression were evaluated using molecular and biochemical assays. Both MCAO/R mice and OGD/R-treated N2a cells exhibited decreased m6A levels and upregulation of lncRNA-AU020206 accompanied by enhanced ferroptosis. METTL3 overexpression increased the m6A modification of AU020206, promoting its degradation and attenuating neuronal injury, whereas silencing AU020206 or overexpressing YTHDC2 decreased SLC7A11 mRNA stability and enhanced ferroptosis. Restoring the expression of SLC7A11/GPX4 can enhance cell viability, alleviate neuronal apoptosis, and reduce Fe2+overload. Disruption of the METTL3–AU020206–YTHDC2 axis abolished these neuroprotective effects. METTL3-mediated m6A modification of lncRNA-AU020206 restrained ferroptosis and neuronal injury in cerebral I/R by maintaining the stability of the SLC7A11/GPX4 axis via interactions with YTHDC2. Targeting this epitranscriptomic signalling pathway may represent a promising therapeutic strategy for the treatment of ischemic stroke and related neurological disorders.

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