Hoechst33342染色液
Hoechst_33342_SDS_MedChemExpress

Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:May-24-2017Print Date:May-24-20171. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :Hoechst 33342Catalog No. :HY-15559CAS No. :23491-52-31.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureNot a hazardous substance or mixture.2.2 GHS Label elements, including precautionary statementsNot a hazardous substance or mixture.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:bisBenzimide H 33342; HOE 33342; Hoechst⁻33342; Hoechst33342Formula:C27H28N6OMolecular Weight:452.55CAS No. :23491-52-34. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature:Powder-20°C 3 years4°C 2 yearsIn solvent-80°C 6 months-20°C 1 monthShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance Off-white to pink (Solid)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGThis substance is considered to be non-hazardous for transport.IATAThis substance is considered to be non-hazardous for transport.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2017 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
细胞染色方法总结

Hoechst染色:hoechst可以穿过活细胞膜与细胞核结合(主要为凋亡活细胞)在紫外光下将核染为蓝色。
Hoechst染细胞核会影响共聚焦显微镜对该样本其他荧光的观察效果。
hoechst有hoechest33342和hoechst33258两种hoechsts33258,hoechst33342二者区别不大,但是hoechst33342对细胞的毒性作用更小一些,所以一般来说hoechsts33258用于细胞固定后再染色,而hoechst33342则可以对活细胞直接进行染色!染色步骤PI(Propidium Iodide碘化丙啶)染色:是一种可对DNA染色的细胞核染色试剂,常用于细胞凋亡检测.碘化丙啶(Propidium Iodide,PI)是一种核酸染料(红色),它不能透过完整的细胞膜,但凋亡中晚期的细胞和坏死细胞由于细胞膜通透性的增加,PI 能够透过细胞膜而使细胞核染红.用PI单一染色观测培养细胞,只能表示细胞的坏死情况,而不是凋亡(当然晚期凋亡PI亦可着色).但是如果您只是想知道细胞的死亡情况,而不是仔细区分坏死或凋亡,那么PI单一染色也可以。
但是如果您一定要认定细胞的凋亡,那么PI单一染色显然不够!annexin—v染色细胞凋亡早期,细胞膜标志发生改变。
其中,磷脂酰丝氨酸(Annexin-V,PS)外翻,Annexin—V 在Ca+存在的条件下与其高亲和力特异性结合。
这样,Annexin-v 染色阳性,表示细胞处于早期凋亡状态。
Annexin—V结合不同的荧光抗体,就可以利用流式细胞仪、荧光显微镜以及共聚焦激光扫描显微镜检测细胞凋亡的发生。
Annexin V用FITC标记发绿色荧光;如果用PE标记就发红色荧光。
JC—1染色JC-1是一种阳离子染料,可以在线粒体内聚集,低浓度时主要以单体(monomer)存在,发射光以绿光(~525nm)为主;而在高浓度时则可以形成多聚体(aggregation),发射光以红光(-590nm)为主。
Hoechst_33342_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:Hoechst 33342 is an AT–specific DNA minor groove ligand used fluorochrome for visualizing cellular DNA .IC50 & Target: Dye reagent [1]DNA Stain [1]In Vitro: Hoechst 33342 binds to adenine–thymine–rich regions of DNA in the minor groove. On binding to DNA, the fluorescence greatly increases. This protocol describes the use of Hoechst 33342 to label nuclear DNA of cells grown in culture. Hoechst 33342can also be used to stain fixed cells by substituting Hoechst 33342 for DAPI [1].PROTOCOL (Extracted from published papers and Only for reference)Cell Assay: Hoechst 33342 (10 mg/mL in H 2O stock solution; protected from light)[1].[1]Labeling Nuclear DNA with Hoechst 33342[1]Step 1, Dilute the Hoechst stock solution 1:100 in H 2O for use in labeling. Step 2, Aspirate the cell medium from cells grown on coverslips. Rinse the cells three times with PBS +. Step 3, Incubate the cells in the Hoechst labeling solution (from Step 1) for 10–30min at room temperature. Step 4, Aspirate the labeling solution. Rinse the cells three times in PBS +. Step 5, Mount the coverslips.Step 6, Image the cells (λex ~353 nm, λem ~483 nm for Hoechst 33342)[1].References:[1]. Chazotte B. Labeling nuclear DNA with hoechst 33342. Cold Spring Harb Protoc. 2011 Jan 1;2011(1):pdb.prot5557.Product Name:Hoechst 33342Cat. No.:HY-15559CAS No.:23491-52-3Molecular Formula:C 27H 28N 6O Molecular Weight:452.55Target:DNA Stain; Autophagy Pathway:Cell Cycle/DNA Damage; Autophagy Solubility:DMSO or H 2OCaution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@ Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
HOCHEST_染色的讨论

Hoechst染色的讨论Hoechst染色的具体步骤caspase8428:我看到资料上说,荧光染色的时候可以用DAB与荧光物质反应生成沉淀。
那么,Hoechst染色时,什么时候加DAB,浓度是多少,用什么稀释,最后怎么来终止反应?是不是还要用抗退色固片介质来封片?sunandsuny:Hoechst染色时无需用DAB来显色,它直接结合细胞的DNA ,经过紫外光激发后发出蓝色荧光。
如果你是用来染培养的细胞的话,可以参考下面的步骤(切片雷同):细胞涂片:甲醇:冰乙酸(3:1)固定15分钟,PBS洗一次,加Hoechest 33258荧光染料37℃孵育15~30分钟,于荧光显微镜下观察。
凋亡细胞由于染色质固缩,细胞核呈致密浓染,或呈碎块状致密浓染。
操作比较容易的。
altbenair:想做肝癌细胞hepG2。
看过很多帖子还有文献上的方法,但是都不太具体大都是这样:固定(福尔马林4%),pbs洗三遍,加hoechst孵育1H,将细胞悬液滴于载玻片上荧光显微镜检测。
细胞悬液怎么做?我要做贴壁细胞用胰酶消化?drake015: .1.贴壁细胞,让细胞自己在盖玻片上爬片。
操作如下:A. 取普通洁净盖玻片于70%乙醇中浸泡5分钟或更长时间,无菌超净台内吹干或用细胞培养PBS 或0.9%NaCl等溶液洗涤三遍,再用细胞培养液洗涤一遍。
将盖玻片置于六孔板内,种入细胞培养过夜,使约为50%-80%满。
B. 刺激细胞发生凋亡后,吸尽培养液,加入0.5ml固定液,固定10分钟或更长时间(可4℃过夜)。
C. 去固定液,用PBS或0.9%NaCl洗两遍,每次3分钟,吸尽液体。
洗涤时宜用摇床,或手动晃动数次。
D. 加入0.5ml Hoechst 33258染色液,染色5分钟。
也宜用摇床,或手动晃动数次。
E. 用PBS或0.9%NaCl洗两遍,每次3分钟。
F. 滴一滴抗荧光淬灭封片液于载玻片上,盖上贴有细胞的盖玻片,尽量避免气泡。
细胞染色方法大全

Hoechst染色:hoechst可以穿过活细胞膜与细胞核结合(主要为凋亡活细胞)在紫外光下将核染为蓝色. Hoechst染细胞核会影响共聚焦显微镜对该样本其他荧光的观察效果.hoechst 有hoechest33342和hoechst33258两种hoechsts33258,hoechst33342二者区别不大,但是hoechst33342对细胞的毒性作用更小一些,所以一般来说hoechsts33258用于细胞固定后再染色,而hoechst33342则可以对活细胞直接进行染色!染色步骤PI (Propidium Iodide碘化丙啶)染色:是一种可对DNA染色的细胞核染色试剂,常用于细胞凋亡检测.碘化丙啶(Propidium Iodide, PI)是一种核酸染料(红色),它不能透过完整的细胞膜,但凋亡中晚期的细胞和坏死细胞由于细胞膜通透性的增加,PI 能够透过细胞膜而使细胞核染红.用PI单一染色观测培养细胞,只能表示细胞的坏死情况,而不是凋亡(当然晚期凋亡PI亦可着色)。
但是如果您只是想知道细胞的死亡情况,而不是仔细区分坏死或凋亡,那么PI单一染色也可以。
但是如果您一定要认定细胞的凋亡,那么PI单一染色显然不够!annexin-v染色细胞凋亡早期,细胞膜标志发生改变.其中,磷脂酰丝氨酸(Annexin-V,PS)外翻,Annexin-V 在Ca+存在的条件下与其高亲和力特异性结合.这样,Annexin-v 染色阳性,表示细胞处于早期凋亡状态.Annexin-V结合不同的荧光抗体,就可以利用流式细胞仪、荧光显微镜以及共聚焦激光扫描显微镜检测细胞凋亡的发生。
Annexin V用FITC标记发绿色荧光;如果用PE标记就发红色荧光。
JC-1染色JC-1是一种阳离子染料,可以在线粒体内聚集,低浓度时主要以单体(monomer)存在,发射光以绿光(~525nm)为主;而在高浓度时则可以形成多聚体(aggregation),发射光以红光(-590nm)为主。
细胞染色方法总结

Hoechst染色:hoechst可以穿过活细胞膜与细胞核结合 (主要为凋亡活细胞)在紫外光下将核染为蓝色. Hoechst染细胞核会影响共聚焦显微镜对该样本其他荧光的观察效果.hoechst有hoechest33342和hoechst33258两种 hoechsts33258,hoechst33342二者区别不大,但是hoechst33342对细胞的毒性作用更小一些,所以一般来说hoechsts33258用于细胞固定后再染色,而hoechst33342则可以对活细胞直接进行染色!染色步骤PI (Propidium Iodide碘化丙啶)染色:是一种可对DNA染色的细胞核染色试剂,常用于细胞凋亡检测. 碘化丙啶(Propidium Iodide, PI)是一种核酸染料(红色),它不能透过完整的细胞膜,但凋亡中晚期的细胞和坏死细胞由于细胞膜通透性的增加,PI 能够透过细胞膜而使细胞核染红.用PI单一染色观测培养细胞,只能表示细胞的坏死情况,而不是凋亡(当然晚期凋亡PI亦可着色)。
但是如果您只是想知道细胞的死亡情况,而不是仔细区分坏死或凋亡,那么PI 单一染色也可以。
但是如果您一定要认定细胞的凋亡,那么PI单一染色显然不够!annexin-v染色细胞凋亡早期,细胞膜标志发生改变.其中,磷脂酰丝氨酸(Annexin-V,PS)外翻 ,Annexin-V 在Ca+存在的条件下与其高亲和力特异性结合.这样,Annexin-v 染色阳性,表示细胞处于早期凋亡状态.Annexin-V结合不同的荧光抗体,就可以利用流式细胞仪、荧光显微镜以及共聚焦激光扫描显微镜检测细胞凋亡的发生。
Annexin V用FITC标记发绿色荧光;如果用PE标记就发红色荧光。
JC-1染色 JC-1是一种阳离子染料,可以在线粒体内聚集,低浓度时主要以单体(monomer)存在,发射光以绿光(~525nm)为主;而在高浓度时则可以形成多聚体(aggregation),发射光以红光(-590nm)为主。
(word完整版)HOCHEST 染色的讨论
Hoechst染色的讨论Hoechst染色的具体步骤caspase8428:我看到资料上说,荧光染色的时候可以用DAB与荧光物质反应生成沉淀。
那么,Hoechst染色时,什么时候加DAB,浓度是多少,用什么稀释,最后怎么来终止反应?是不是还要用抗退色固片介质来封片? sunandsuny:Hoechst染色时无需用DAB来显色,它直接结合细胞的DNA ,经过紫外光激发后发出蓝色荧光。
如果你是用来染培养的细胞的话,可以参考下面的步骤(切片雷同):细胞涂片:甲醇:冰乙酸(3:1)固定15分钟,PBS洗一次,加Hoechest 33258荧光染料37℃孵育15~30分钟,于荧光显微镜下观察。
凋亡细胞由于染色质固缩,细胞核呈致密浓染,或呈碎块状致密浓染.操作比较容易的。
altbenair:想做肝癌细胞 hepG2。
看过很多帖子还有文献上的方法,但是都不太具体大都是这样:固定(福尔马林4%),pbs洗三遍,加hoechst孵育1H,将细胞悬液滴于载玻片上荧光显微镜检测.细胞悬液怎么做?我要做贴壁细胞用胰酶消化?drake015:。
1.贴壁细胞,让细胞自己在盖玻片上爬片。
操作如下:A. 取普通洁净盖玻片于70%乙醇中浸泡5分钟或更长时间,无菌超净台内吹干或用细胞培养PBS或0。
9%NaCl等溶液洗涤三遍,再用细胞培养液洗涤一遍。
将盖玻片置于六孔板内,种入细胞培养过夜,使约为50%—80%满。
B. 刺激细胞发生凋亡后,吸尽培养液,加入0。
5ml固定液,固定10分钟或更长时间(可4℃过夜)。
C。
去固定液,用PBS或0.9%NaCl洗两遍,每次3分钟,吸尽液体。
洗涤时宜用摇床,或手动晃动数次。
D. 加入0。
5ml Hoechst 33258染色液,染色5分钟.也宜用摇床,或手动晃动数次。
E. 用PBS或0。
9%NaCl洗两遍,每次3分钟.F。
滴一滴抗荧光淬灭封片液于载玻片上,盖上贴有细胞的盖玻片,尽量避免气泡。
使细胞接触封片液,切勿弄反。
常见细胞核荧光染料
细胞核常用荧光染料有:吖啶橙(Acridine Orange,AO)、溴化乙锭(Ethidium Bromide,EB)和碘化丙啶(Propidium Iodide,PI),DAPI、Hoechst染料、EthD III、7-AAD、RedDot1、2 等等。
透膜的染料如下:AO:具有膜通透性,能透过细胞膜,将核DNA和RNA分别染成绿色和红色,因此使细胞核呈绿色或黄绿色荧光。
EB:一种高度灵敏的荧光染色剂,在标准302nm处激发出橙红色信号。
DAPI:蓝色一种可以穿透细胞膜的蓝色荧光染料,其与DNA结合后可以产生比DAPI自身强20多倍的荧光,而与单链DNA结合无荧光的增强。
DAPI对双链DNA 的染色灵敏度要高于EB和PI,荧光强度比Hoechst低,但光稳定性高于Hoechst。
Hoechst染料:蓝色一类在显微观察中标记DNA的荧光染料,最常见的两种是Hoechst33342和Hoechst33258。
这两种染料都在紫外350nm处被激发,在461nm 处最大发射光附近发射青/蓝色荧光。
与DAPI相比,Hoechst33342加有乙基,具有更强的亲脂性,因此能更好的透过完整的细胞膜,并且细胞毒性更小。
RedDot 1染料:红色,超强的细胞核选择性,其光谱相似于Draq?5 和 Draq?7。
RedDot?染料可被几种常见的激光激发并可在远红外区激发荧光。
RedDot? 的红色近红外荧光有效的与其他常用荧光探针区分开来。
不透膜的染料,如下:PI:不同通过活细胞膜,但却能穿过破损的细胞膜而对核染色。
PI作为红色荧光复染剂首选,PI经常与Calcein-AM或者FDA等荧光探针合用,区分死/活细胞。
EthD III、7-AAD、RedDot 2:不能透过细胞膜,但能将坏死细胞区分开来;更适合凋亡坏死实验的检测;细胞核荧光染料(PI DAPI Hoechst33342)细胞核荧光染料PI碘化丙啶(简称PI)是一种常用的细胞核荧光染色剂。
Hoechst 33342_23491-52-3_DataSheet_MedChemExpress
Product Name:Hoechst 33342CAS No.:23491-52-3Cat. No.:HY-15559Product Data SheetMWt:452.55Formula:C27H28N6O Purity :>98%Solubility:DMSO or water Protect from lightMechanisms:Biological Activity:Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA Hoechst 33342is a Pathways:Others; Target:DNA Stain Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA. Hoechst 33342 is acell dye for DNA.IC50 Value: These Bis-benzimides were originally developed by Hoechst AG, which numbered all theircompounds so that the dye Hoechst 33342 is the 33342nd compound made by the company. There are three related Hoechst stains: Hoechst 33258, Hoechst 33342, and Hoechst 34580. The dyes Hoechst 33258 and Hoechst 33342 are the ones most commonly used and they havesimilarexcitation/emission spectra. Both dyes are excited by ultraviolet light at around 350 nm, and both emit blue/cyan fluorescent light around anemission maximum at 461 nm. Unbound dye has its References:[1]. Latt SA, Stetten G, Juergens LA, Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence. The journal of histochemistry and cytochemistry : officialjournal of the Histochemistry Society 23 (7): 493-505.maximum fluorescence emission in the 510-540 nm range. Hoechst dyes are soluble in water and in organic solvents such as dimethyl formamide or dimethyl sulfoxide. Concentrations can be achieved of up to 10 m...j y y ()[2]. a b c "Hoechst Stains". Invitrogren (Molecular Probes).[3]. Portugal J, Waring MJ. Assignment of DNA binding sites for 4',6-diamidine-2-phenylindole and bisbenzimide (Hoechst 33258). A comparative footprinting study. Biochimica et Biophysica Acta 949(2): 158-68.Caution: Not fully tested. For research purposes onlyMedchemexpress LLC18W i l k i n s o n W a y , P r i n c e t o n , N J 08540,U S AE m a i l : i n f o @m e d c h e m e x p r e s s .c o m W e b : w w w .m e d c h e m e x p r e s s .c o m。
DAPI染色液使用说明书
DAPI染色液使用说明书DAPI染色液(DAPI Staining Solution)是经过精心优化几乎适用于所有常见细胞和组织细胞核染色的染色液。
DAPI,即2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochlorid e,也称DAPI dihydrochloride,分子式为C16H15N5·2HCl ,分子量为350.25 ,CAS Number28718-90-3。
DAPI是一种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。
和EB(ethidiumbromide)相比,对双链DNA的染色灵敏度要高很多倍。
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。
本DAPI染色液可以直接用于固定细胞或组织的细胞核染色。
包装清单:保存条件:-20℃避光保存,一年有效。
注意事项:本DAPI染色液的浓度经过碧云天的优化,确保可以满足各种常规染色的需要。
如需使用特定浓度的DAPI,请选购碧云天的DAPI(C1002)。
荧光染料都存在淬灭的问题,建议染色后尽量当天完成检测。
为减缓荧光淬灭可以使用抗荧光淬灭封片液。
抗荧光淬灭封片液(P0126)可以向碧云天订购。
DAPI对人体有刺激性,操作时请小心,并注意适当防护以避免直接接触人体或吸入体内。
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。
为了您的安全和健康,请穿实验服并戴一次性手套操作。
使用说明:1.对于细胞或组织样品,固定后,适当洗涤去除固定剂。
随后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染色。
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Hoechst 33342染色液
产品简介:
Hoechst 33342也称bisBenzimide H 33342或HOE 33342,分子式为C27H28N6O·3HCI·3H2 O,分子量为615.99,CAS Number 23491-52-3. Hoechst 33342是一种可以穿透细胞膜的蓝色荧光染料,对细胞的毒性较低,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。
Hoechst 33342也用于普通的细胞染色、DNA染色。
Hoechst 33342的最大激发波长为346nm,最大发射波长为460 nm。
Hoechst 33342和双链DNA结合后,最大激发波长为352 nm,最大发射波长为461 nm。
Jimei Hoechst 33342染色液直接用于固定细胞或组织的细胞核染色,也可直接用于活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):
(一)固定的组织细胞染色
1、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Hoechst 33342染色,如果
不需要进行其它染色,则直接进行后续的Hoechst 33342染色。
对于贴壁细胞或
组织切片,加入少量Hoechst 33342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Hoechst 33342染色工作液,充分混匀。
2、室温放置5~8min。
3、轻轻吸除Hoechst 33342染色液。
4、用无菌的PBS或生理盐水清洗2~3次,每次3~5min。
5、直接在荧光显微镜下观察或封片后荧光显微镜下观察。
(二)活细胞染色
1、取96、24、6孔板培养细胞至合适状态,按96孔板加入100μl、24孔板加入500
μl、6孔板加入1ml的比例,加入适当的Hoechst 33342染色工作液,染液必须
充分覆盖细胞。
2、在适宜于细胞培养的条件下培养20~30 min。
3、轻轻吸除Hoechst 33342染色液。
4、用无菌的PBS或生理盐水清洗2~3次,每次3~5min。
5、进行荧光检测。
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、Hoechst 33342染色液的浓度适用于各种常规染色的需要。
2、荧光染料都存在淬灭的问题,建议染色后尽快检测。
3、为减缓荧光淬灭,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、Hoechst 33342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,请穿实验服并戴一次性手套操作。
有效期:6个月有效。
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