Caspase_1抑制剂_激动剂_MCE
Caspase活性检测说明书
Caspase活性检测（基于p NA标记底物的比色法）在caspase 的底物多肽上标记发色团如：p NA，可用于检测凋亡细胞中的caspase活性及筛选caspases的激活剂或抑制剂。
本操作步骤主要针对使用酶标仪进行读数而设计。
其中使用的细胞抽提物、底物、抑制剂及p NA的用量为每个样本140ul。
若您希望设计针对分光光度计读数的实验，建议对每种溶液体积进行进一步优化。
所需溶液、试剂及设备• Caspase底物• Caspase抑制剂• 细胞裂解缓冲液：50mM HEPES, pH7.4, 100mM NaCl, 0.1%CHAPS, 1mM DTT, 100uM EDTA • 检测缓冲液：50mM HEPES, pH7.4, 100mM NaCl, 0.1%CHAPS, 10mM DTT, 100uM EDTA, 10%Glycerol• PBS：将8g NaCl、200mg KCl和1.44g Na2HPO4溶于800ml蒸馏水；用HCl调至pH7.4；定容至1L• 酶标板和酶标仪其它试剂(推荐)• 纯化的Caspase（阳性对照）• p NA细胞抽提物的制备以适合的凋亡诱导剂对细胞进行诱导（参考前述操作方法），同时做阴性对照，包括未处理细胞、用诱导剂的无诱导活性类似物处理细胞（若可得到），或一个凋亡诱导处理“零时间”样品。
应保证有足够细胞进行重复实验，包含或不含抑制剂的对照以检测蛋白浓度。
一般来说，我们下面推荐的裂解前细胞数量可以产生足够蛋白浓度供酶标仪检测；然而不同大小、体积的细胞及蛋白浓度可能需要增加铺板密度。
当裂解2×107/ml细胞是，产生的蛋白浓度约为1-3mg/ml，体积10ul，含约10-30ug蛋白。
依据其细胞系，最少应有106个细胞，体积50ul的裂解缓冲液进行处理。
• 细胞计数，离心收集细胞。
以PBS缓冲液洗涤一次。
若细胞已被caspase抑制剂处理过，建议多洗几次以防止后继实验中可能造成的不利影响。
MedChemExpress抑制剂Cocktail家族全系列产品,为您的蛋白质检测保驾护航!
MCE抑制剂Cocktail家族全系列产品，为您的蛋白质检测保驾护航！“曾经，有一批待检蛋白摆在我的面前，我没有及时检测，等到它降解了，我才后悔莫及。
实验中最痛苦的事莫过于此……只能，再提一批！”实验室的故事，说多了都是泪啊。
尤其蛋白质的研究，一不小心样品就降解了、去乙酰化了，检测结果必然一无所获。
还好有MCE inhibitor cocktail家族为您的蛋白质提供全方位的保护。
蛋白酶抑制剂、磷酸酶抑制剂、去乙酰化酶抑制剂……不管您的课题是细胞通路、肿瘤研究、蛋白组学研究、免疫研究等，总有一款适合您！快点击以下产品链接，申请免费试用吧！MCE抑制剂产品介绍HY-K0010Protease Inhibitor Cocktail (EDTA-Free, 100X in DMSO)用于细胞裂解与组织抽提。
HY-K0011Protease Inhibitor Cocktail, mini-Tablet (EDTA-Free)用于细胞裂解与组织抽提，片剂更便于使用。
HY-K0021Phosphatase Inhibitor Cocktail I (100X in DMSO)有效抑制碱性、丝氨酸/苏氨酸磷酸酶。
HY-K0022Phosphatase Inhibitor Cocktail II (100X in ddH2O)有效抑制酸性、碱性、酪氨酸磷酸酶。
HY-K0023Phosphatase Inhibitor Cocktail III (100X in DMSO)有效抑制碱性、丝氨酸/苏氨酸磷酸酶。
HY-K0030Deacetylase Inhibitor Cocktail (100X in 70% DMSO)有效抑制蛋白的去乙酰化作用。
*试用装详情请咨询销售。
HY-16658_Z-VAD-FMK_MCE
Inhibitors, Agonists, Screening LibrariesData SheetBIOLOGICAL ACTIVITY:Z–VAD–FMK is a cell–permeable, pan–caspase inhibitor.IC50 & Target: pan–caspase [1]In Vitro: Z–VAD–FMK is a broad–spectrum caspase inhibitor, has been shown to inhibit the intracellular activation of caspase–likeproteases. The injection of Z–VAD–FMK suppresses the caspase–3 activity in lung tissues, and significantly decreases the number of terminal dUTP nick–end labeling–positive cells [1]. Z–VAD–FMK is administered intraperitoneally at 1 hour before and 6 hours after SAH. Expression of caspase–3 and positive TUNEL is examined as markers for apoptosis. Z–VAD–FMK suppresses TUNEL and caspase–3 staining in endothelial cells, decreases caspase–3 activation, reduces BBB permeability, relieves vasospasm, abolishes brain edema, and improves neurological outcome [2]. Z–VAD–FMK is a cell–permeable caspase inhibitor, efficiently blocks cell deathinduced by SMN deficiency [3].In Vivo: The survival rate of mice is prolonged significantly by the injection of Z–VAD–FMK. All mice succumbed to LPS within 30hours. By contrast, the mice treated with Z–VAD–FMK survive significantly longer and 27% of the mice survived more than 7 days [1].PROTOCOL (Extracted from published papers and Only for reference)Cell Assay: Z–VAD–FMK is dissolved in DMSO and stored, and then diluted with appropriate medium before use [3]. [3]PCR products containing coding sequences for the dSMN (forward primer: 5′–TAA TAC GAC TCA CTA TAG GG AAG ACG TAC GAC GAG TCG–3′; and reverse primer: 5′–TAA TAC GAC TCA CTA TAG GG GTG GTG CTG GCT TCT TTC–3′; product length, 601bps; bold and italics letters represent T7 promoter sequences) and control Drosophila Presenilin (dPsn) gene (forward primer: 5′–TAA TAC GAC TCA CTA TAG GG TG GCT GCT GTC AAT CTC–3′; and reverse primer: 5′–TAA TAC GAC TCA CTA TAG GG CGA TAG CAA CGC TTC TTG–3′; product length: 543bps) are obtained and gel–purified. Double–stranded RNAs (dsRNA) are generated by transcription with Ribomax T7Transcription kit and digested with Rnase–free DNase. The dsRNA products are ethanol precipitated and annealed by incubation at 65°C for 30 min and then slowly allowed to cool at room temperature. The annealed dsRNA products are analyzed on a 1% agaorse gel to ensure the majority of dsRNA existed as a single band. The dsRNA (2 μg) and/or plasmid DNAs (2 μg) are introduced into cells by using Cellfectin. Caspase inhibition is achieved by using 50 μM of Z–VAD–FMK in the culture medium [3].Animal Administration: Z–VAD–FMK is dissolved at 2 mg/mL in 1% DMSO in sterile saline (Mice)[1].Z–VAD–FMK is dissolved in 1% DMSO and further diluted in physiological buffer solution (final <0.01% DMSO) (Rat)[2].[1][2]Mice [1]Mice used in this study are 5– to 6–week–old (20 to 22 g) ICR males. Mice are injected with 30 mg/kg LPS from E. coli serotype O111:B4 through the tail vein. A single intravenous injection of Z–VAD.fmk (0.25 mg) is made 15 minutes before LPS injection,followed by three intravenous injections of Z–VAD–FMK (0.1 mg each) per hour. Control mice are injected with the same volume of 1%DMSO in sterile saline.Rat [2]Product Name:Z–VAD–FMK Cat. No.:HY-16658CAS No.:187389-52-2Molecular Formula:C 22H 30FN 3O 7Molecular Weight:467.49Target:Caspase Pathway:ApoptosisSolubility:DMSO: ≥ 30 mg/mLMale Sprague–Dawley rats weighing 300 to 350 g are anesthetized with α–chloralose (40 mg/kg IP) and urethane (400 mg/kg IP). Animals are intubated, and respiration is maintained with a small animal respirator. Rectal temperature is maintained at 37±0.5°C with a heating pad. The left external carotid artery is isolated and a 4.0 monofilament nylon suture is inserted through the internal carotid artery to perforate the middle cerebral artery. SAH is confirmed at autopsy in each rat. Sham–operated rats underwent the same procedures except that the suture is withdrawn after resistance is felt. Z–VAD–FMK (50 μM per 0.3 mL) is injected intraperitoneally at 1 hour before and 6 hours after SAH induction. In vehicle group, rats underwent SAH induction and are treated with the same volume of vehicle (DMSO diluted in physiological buffer solution). No treatment is applied in sham–operated animals.References:[1]. Kawasaki M, et al. Protection from lethal apoptosis in lipopolysaccharide–induced acute lung injury in mice by a caspaseinhibitor. Am J Pathol. 2000 Aug; 157(2):597–603.[2]. Park S, et al. Neurovascular protection reduces early brain injury after subarachnoid hemorrhage. Stroke. 2004 Oct;35(10):2412–7.[3]. Ilangovan R, et al. Inhibition of apoptosis by Z–VAD–fmk in SMN–depleted S2 cells. J Biol Chem. 2003 Aug 15;278(33):30993–9.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, USACertificate of AnalysisPHYSICAL AND CHEMICAL PROPERTIESMolecular Formula:C 22H 30FN 3O 7Molecular Weight:467.49Storage:Powder -20°C 3 years 4°C 2 years In solvent-80°C 6 months -20°C1 monthChemical Structure:ANALYTICAL DATAAppearance:White to off-white (Solid)1H NMR Spectrum:Consistent with structure Purity (NMR):>98.0%Conclusion:The product has been tested and complies with the given specifications.Product Name:Z-VAD-FMK Cat. No.:HY-16658CAS No.:187389-52-2Batch No.:25378Chemical Name:L-Alaninamide, N-[(phenylmethoxy)carbonyl]-L-valyl-N-[(1S)-3-fluoro-1-(2-methoxy-2-oxoethyl)-2-oxopropyl]-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, USASafety Data Sheet Revision Date:Jul.-17-2017Print Date:Jul.-17-20171. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :Z-VAD-FMKCatalog No. :HY-16658CAS No. :187389-52-21.2 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:Z–VAD(OMe)–FMK; Z–Val–Ala–Asp(OMe)–FMKFormula:C22H30FN3O7Molecular Weight:467.49CAS No. :187389-52-24. 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 White to off-white (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, USAppm (t1)0.01.02.03.04.05.06.07.08.09.010.08.6368.6178.4898.4698.3808.3738.3608.1908.1758.1408.1267.4267.4057.3927.3607.3327.3267.3107.2885.2895.2515.2355.2095.1705.1345.1185.0915.0274.6544.6404.6294.2114.1934.1713.8983.8833.8653.5983.3412.8592.8412.8352.8172.8002.6822.6752.6692.6492.6322.6102.5912.5072.4412.3332.0691.9711.9551.9391.9221.9141.2401.2051190871564508295.9753.8551.0001.0231.0343.0911.0501.1091.0333.0546.0982.048Date:14 Jul 2017Document's Title:Catalog No: HY-16658 Batch#25378Spectrum Title:biz2017-714-wyq17-dmso-170714Frequency (MHz):(f1) 400.130Original Points Count:(f1) 32768Actual Points Count:(f1) 32768Acquisition Time (sec):(f1) 3.2768Spectral Width (ppm):(f1) 24.992 Pulse Program:UnknownSample ID: BIZ2017-714-WYQ17, Catalog No: HY-16658 Batch#25378 , DMSON HOO OO F H NO N HOO Chemical Formula:C 22H 30FN 3O 7Exact Mass:467.21。
mce铁死亡脂质过氧化实验步骤
mce铁死亡脂质过氧化实验步骤引言：mce（Malondialdehyde and Cholesterol Ester）铁死亡脂质过氧化实验是一种常用的体外实验方法，用于评估细胞膜脂质过氧化程度。
本文将详细介绍该实验的步骤及操作流程。
一、实验前准备1. 准备所需试剂和设备，包括mce试剂盒、离心机、超声波清洗器等。
2. 温度控制：将实验室温度控制在37摄氏度。
二、样品准备1. 收集需要进行实验的样品，如细胞培养物或动物组织。
2. 将样品放入离心管中，并加入PBS缓冲液悬浮均匀。
3. 离心样品，去掉上清液。
三、实验操作1. 向样品中加入mce试剂，按比例加入适量的试剂。
2. 轻轻振摇样品，使试剂充分混合。
3. 将样品置于37摄氏度的恒温水浴中孵育一定时间，一般为30分钟。
4. 取出样品，离心使沉淀物沉积在离心管底部。
5. 将上清液转移到新的离心管中。
6. 加入适量的氯仿，并轻轻振摇混合。
7. 离心样品，使沉淀物和氯仿分离。
8. 取上清液转移到新的离心管中。
9. 加入适量的PBS缓冲液，并轻轻振摇混合。
10. 离心样品，使沉淀物和PBS缓冲液分离。
11. 取上清液转移到新的离心管中。
四、测量结果1. 取出上清液，加入相应试剂。
2. 按照试剂盒说明书的要求，进行相应的检测，如比色法、荧光法等。
3. 根据试剂盒提供的标准曲线，计算样品中的mce含量。
五、数据分析1. 对测量结果进行统计学分析，计算平均值和标准差。
2. 根据实验设计和目的，对数据进行图表展示和解释。
3. 进一步分析结果，探讨实验的意义和可能的影响因素。
六、实验注意事项1. 实验操作过程中需严格遵守无菌操作规范，避免污染。
2. 每一步操作都要准确无误，避免操作失误对实验结果的影响。
3. 在实验过程中，注意安全操作，避免对人身及环境造成危害。
4. 仪器设备的使用和维护要规范，确保实验的准确性和可重复性。
结论：mce铁死亡脂质过氧化实验是一种简单有效的体外实验方法，用于评估细胞膜脂质过氧化程度。
DNA_methyltransferase_(DNA_MTase)抑制剂_激动剂_MCE
DNMTDNA methyltransferase (DNA MTase)the transfer of a methyl group to DNA. DNA methylation serves awide variety of biological functions. All the known DNAmethyltransferases use S-adenosyl methionine (SAM) as the methyldonor. MTases can be divided into three different groups on the basisof the chemical reactions they catalyze:m6A, m4C, m5C. m6A andm4C methyltransferases are found primarily in prokaryotes. m5Cmethyltransfereases are found in some lower eukaryotes, in mosthigher plants, and in animals beginning with the echinoderms. Threeactive DNA methyltransferases have been identified in mammals.They are DNMT1, DNMT3A and DNMT3B. A fourth enzyme previouslyknown as DNMT2 is not a DNA methyltransferase.DNMT Inhibitors & ModulatorsDecitabine is a potent inhibitor of DNA methyltransferase with IC50of 438 nM and 4.38 nM in HL-60 and KG1a cells, respectively.O6BTG-C8-(beta)Glu is a glucose-conjugated MGMT inhibitor with IC50 values of 32 nM in vitro and 10 nM on HeLa S3 cell; the parent Lomeguatrib (PaTrin-2) is a modified guanine base, which can repress the activity of DNA repair protein O(6)-alkylguanine-DNA Procainamide, it is a Class III antiarrhythmic agent, whereas RG108 is a non-nucleoside inhibitor of DNA methyltransferase with SGI-1027 is a potent and selective inhibitor of DNA methyltransferase (DNMT) with IC50 of 6, 8, 7.5 μM for DNMT1, DNMT3A, and Zebularine(NSC309132; 4-Deoxyuridine) is a DNA methylationinhibitor that forms a covalent complex with DNA methyltransferases,Email: sales@Cat. No.: HY-A0004Cat. No.: HY-13057Cat. No.: HY-13668Cat. No.: HY-B1109Cat. No.: HY-13642Cat. No.: HY-13962Cat. No.: HY-13420。
csapase分子量
csapase分子量摘要：一、CASPASE酶简介二、CASPASE酶的分子量三、CASPASE酶在生物学中的应用四、CASPASE酶的研究意义五、总结正文：【一、CASPASE酶简介】CASPASE（Cysteine Aspartate-Specific Protease）是一种半胱氨酸天冬氨酸特异性蛋白酶，广泛存在于生物体内。
它是一类重要的凋亡调控因子，能够诱导细胞凋亡，参与多种生物过程。
【二、CASPASE酶的分子量】CASPASE酶的分子量因物种和亚型而异。
在人源CASPASE酶中，CASPASE-1、CASPASE-2、CASPASE-4、CASPASE-5的分子量约为35-40kDa，而CASPASE-3、CASPASE-6、CASPASE-7、CASPASE-8、CASPASE-9的分子量约为40-50kDa。
在鼠源CASPASE酶中，分子量也存在类似差异。
【三、CASPASE酶在生物学中的应用】CASPASE酶在生物学中的应用广泛，主要包括：1.细胞凋亡调控：CASPASE酶是细胞凋亡信号通路中的关键执行者，通过激活下游底物蛋白，引发细胞凋亡。
2.免疫调节：CASPASE酶在免疫细胞活化、免疫应答过程中发挥重要作用，如CASPASE-1参与炎症反应。
3.神经系统疾病：CASPASE酶在神经元凋亡过程中发挥作用，与神经系统疾病的发生发展密切相关。
4.肿瘤治疗：CASPASE酶可作为肿瘤治疗的靶点，通过诱导肿瘤细胞凋亡达到治疗目的。
【四、CASPASE酶的研究意义】CASPASE酶的研究意义在于：1.有助于深入了解细胞凋亡及其调控机制，揭示生命现象的本质。
2.为治疗相关疾病提供新思路，如通过调控CASPASE酶活性治疗肿瘤、神经系统疾病等。
3.为生物科学研究提供重要工具，如CASPASE酶抑制剂可用于研究细胞凋亡信号通路。
【五、总结】CASPASE酶是一类重要的半胱氨酸天冬氨酸特异性蛋白酶，具有调控细胞凋亡、免疫反应等多种生物过程的功能。
z-vad原理
Z-V AD原理
Z-V AD-FMK（Zinc-Binding Caspase-inhibitory Peptide with a Fluorescent Marker）是一种细胞渗透性、不可逆的半胱氨酸蛋白酶抑制剂，能够特异性地结合caspase酶，阻止其活化，进而抑制凋亡的发生。
其工作原理如下：
1. Z-V AD-FMK能够与caspase酶的活性中心结合，形成一个稳定的酶-抑制剂复合物。
这种复合物可以防止caspase 酶的切割活化，从而抑制凋亡的发生。
2. Z-V AD-FMK具有荧光标记，可以通过荧光显微镜或流式细胞仪等技术进行检测，实现对细胞凋亡进程的实时监测。
3. Z-V AD-FMK是一种广谱的caspase抑制剂，可以抑制caspase家族的多个成员，如caspase-1、caspase-2、caspase-3、caspase-6和caspase-7等。
4. Z-V AD-FMK的使用可以通过阻断caspase途径来减轻细胞应激，从而保护细胞免受损伤。
总的来说，Z-V AD-FMK通过与caspase酶的活性中心结合，阻止其活化，进而抑制凋亡的发生，并且具有荧光标记，可以实现对细胞凋亡进程的实时监测。
Caspase
Caspase生化特性caspase家族属于半胱氨酸蛋白酶，一般在细胞内以无活性酶形式存在；当其作用于特异性底物，产生异二聚体，同时发出光子。
Caspase检测凋亡的原理通过检测细胞内caspase酶活性了解细胞的凋亡功能。
试剂渗透进入细胞后，使细胞内caspase 酶释放，作用于其特异性底物，产生发光体或荧光，发光酶（荧光酶）作用于发光体，产生光。
发光度与细胞内caspase酶活性成正比。
Caspase酶活性反映了细胞内凋亡途径激活情况，即细胞的凋亡能力。
如何做对照组？阴性对照：caspase抑制剂和培养基；或者只加完全培养基。
试剂盒没有caspase抑制剂？不必要。
Caspase试剂盒组成：caspase3/7特异性底物；含发光酶，细胞溶解液的缓冲液。
Caspase试剂盒分类：caspase3/7,caspase8,caspase9.(后两个是内源性凋亡途径参与物）试剂盒的选择：一般细胞凋亡检测选用caspase3/7试剂盒，如果用于分析凋亡上游启动子，根据对凋亡途径（内源性/外源性）的兴趣，选择caspase8/caspase9试剂盒即可。
Caspase3/7介绍Caspase3/7是一种发光分析法，可用于检测粘附细胞或悬浮生长细胞内caspase3/7的酶活性。
试剂盒中包含了发光前体caspase3/7的Z-DEVD-aminoluciferin 冻干底物，DEVD为抑制剂，保证了试剂盒的特异性。
底物被切割后产生发光体，发光体作为发光酶的底物与之反应后，发出光。
Caspase3/7试剂已经对caspase酶活力，发光酶活力以及细胞溶解做了优化。
将试剂以混合物形式加入样本，导致细胞溶解，底物的切割和产生流性光信号。
可用于研究细胞凋亡和凋亡抑制。
实验大概需用2小时一轮。
检测波长一般检测发光体没有内置的波长选择功能，都采用全部可见光检测。
原因有二：1.不需用滤光；2.滤光后敏感度降低。
内源性凋亡途径：病毒，紫外线或线粒体胞膜破坏，细胞色素C释放至胞液等因素导致内源性凋亡途径激活，从而产生含有caspase-9前体的凋亡小体复合物，复合物导致caspase-9活化，接着caspase-9使下游介导凋亡的死亡效应caspase酶活化，包括caspase3,caspase6,caspase7。