A 77-01_607737-87-1_DataSheet_MedChemExpress
- 格式:pdf
- 大小:100.85 KB
- 文档页数:1


Endotoxin Removal Solution Catalog Number E4274Product DescriptionEndotoxins are lipopolysaccharides (LPS), a major component of the Gram-negative bacterial cell wall, and are commonly found as contaminants in plasmid DNA preparations from E. coli. Endotoxins are large, negatively charged molecules that co-purify with DNA on ion exchange and size exclusion columns and in CsCl banding. Endotoxins are extremely potent stimulators of the mammalian immune system and are toxic to primary cells and to animals. The endotoxin toxicity is an obstacle to in vitro and in vivo transfection experiments.Non-ionic detergents, traditionally used for separation of integral membrane proteins,1 can be utilized for removal of endotoxins from DNA solutions by phase separation.2The solubility behavior of a detergent in a dilute, aqueous solution at physiological salt and pH conditions is strongly dependent upon the temperature of the solution. At low temperatures, the detergent forms a clear, micellar solution, but above the cloud point temperature, the micelles form larger, turbid aggregates and ultimately fuse to form a separate phase. The lower phase is detergent-enriched and the detergent-depleted upper phase contains detergent at a concentration slightly above the critical micellar concentration (CMC). Amphiphilic and hydrophobic molecules associated with the micelles of the detergent will aggregate within the detergent-enriched phase, while the soluble, hydrophilic molecules will remain in the detergent-depleted upper phase.Extraction of endotoxin contaminated DNA solutions with the appropriate non-ionic detergent will separate the hydrophilic DNA from the amphiphilic endotoxin. The amphiphilic endotoxin will associate with the lower phase, while the DNA will remain in the upper, detergent-depleted phase.2Reagents and equipment required, but not provided • Water, Molecular Biology Reagent, Catalog Number W4502• E-TOXATE® Water, Catalog Number 2107, or Tris-EDTA (TE) buffer 100×, Catalog NumberT9285• DNA solution (0.5 ml), ~ 1 mg/ml in E-TOXATE®Water or TE buffer• 3 M sodium acetate solution, pH 7.5.• 2-Propanol, Catalog Number I9516, or Ethanol, 190 proof, Catalog Number E7148; 200 proof, CatalogNumber E7023• 70% Ethanol• E-TOXATE®reagents Kits, Catalog Numbers 210A1, 210B1 or 210C1• Ice bucket• Heat block or incubator at 37 °C• Microcentrifuge at room temperature• 1.5 or 2 ml sterile microcentrifuge tubes• Endotoxin-free pipet tips (40-200 µl, 200-1000 µl) Precautions and DisclaimerThis product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.StorageStore at room temperature.Note: Removal of endotoxins from DNA preparations can be performed either during the final stage of DNApreparation, or during an earlier stage.Procedures for Endotoxin RemovalDuring the final stage of DNA preparationNote: The procedure described below was performed on plasmid DNA produced in E. coli DH5α cells.• Losses of up to 50% of the DNA are expected. • Use of a DNA concentration above therecommended 1 mg/ml reduces the efficiency ofthe procedure.1. Pipette 500 µl of the DNA solution into a sterilemicrocentrifuge tube.2. Add 50 µl of the 3 M sodium acetate solution to theDNA sample.3. Incubate on ice for 5 minutes.4. Add 100 µl of cold Endotoxin Removal Solution.5. Mix thoroughly and incubate on ice for 10 minutes.The solution should be light blue and clear.6. Incubate the tube at 37 °C for 20 to 30 minutes oruntil the phases separate.7. Spin for 5 minutes at 3000 x g in themicrocentrifuge. The upper phase is colorless and clear, while the lower phase is blue.8. Carefully transfer the upper phase containing theDNA to a clean microcentrifuge tube.9. Repeat steps 4 through 8 twice.10. Add 0.6× volume of 2-propanol. Mix by inversion atroom temperature and centrifuge at 15,000 x g for30 minutes at 4 °C. Alternatively, add2.5× volumes of ethanol. Incubate overnight at –20°C or 20 minutes at –70 °C and centrifuge at15,000 x g for 30 minutes at 4 °C.11. Carefully remove the supernatant12. Wash the DNA pellet twice with cold 70% ethanol.Remove the supernatant.13. Air-dry the pellet.14. Suspend the DNA in 100 µl of endotoxin free wateror TE buffer.15. Determine DNA concentration and endotoxin levelsusing endotoxin assay reagents and compare tothe starting material. During an earlier stage of DNA preparationThis procedure is based on the alkaline lysis of E. coli DH5α cells.3 The endotoxins are removed immediately after alkaline cell lysis, neutralization, and a clarification step. The resulting high salt solution is suitable for the endotoxin removal step. It is performed under “endotoxin free” conditions. The plasticware used is either sterile and disposable, or NaOH-treated. The buffers are prepared with endotoxin free water.1. Add the Endotoxin Removal Solution (0.2× volume)to the cold, crude DNA solution.2. Incubate on ice and mix occasionally by inversionto obtain a homogenous, clear blue solution3. Incubate at 37 °C for 20 to 30 minutes until thephase separation is obvious.4. Spin for 5 minutes at low speed (3000 x g) at roomtemperature.5. Transfer the upper aqueous phase to an endotoxinfree container.6. Proceed with the DNA purification by any method.Use endotoxin-free buffers and containers. References1. Bordier, C., J. Biol. Chem., 256, 1604-1607, (1981).2. Cotten, M. et al., Gene Therapy, 1, 239-246,(1994).3. Sambrook et al., Molecular Cloning, a LaboratoryManual, 2nd Ed. p. 1.38RK,PHC 09/05-1Sigma brand products are sold through Sigma-Aldrich, Inc.Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side ofthe invoice or packing slip.。
Inhibitors, Agonists, Screening Libraries Data SheetBIOLOGICAL ACTIVITY:DMP 777 is a potent, selective, and orally active human leukocyte elastase (HLE) inhibitor.In Vivo: DMP–777–treated rats show a marked decrease in H/K–ATPase staining parietal cells. DMP–777–induced loss of parietal cells is significantly ameliorated with coadministration of omeprazole. DMP–777–treated animals demonstrates marked foveolar hyperplasia in the fundus with prominent expansion of diastase–resistant, PAS–positive surface mucous cells. When DMP–777 is coadministered with omprazole, there is a significant decrease in BrdUpositive S–phase cells compared with rats thatreceive DMP–777 alone [1]. After oral dosing of monkeys at 40 mg/kg with DMP–777 the only stereoisomer detected in the post–dose plasma samples is the starting material DMP–777, and no inversion of the configuration at positions 'a' and 'b' of DMP–777 has occurred in vivo [2]. Mist1–/– mice treated with DMP–777 show fewer chief cell to SPEM transitions. Mist1–/– mice treated with L635demonstrates significantly fewer proliferative SPEM cells compared to control mice [3]. PROTOCOL (Extracted from published papers and Only for reference)Animal Administration: DMP–777 is formulated as a suspension in 0.5% methylcellulose. [1]Groups 1A and 1B receive control vehicle instead of omeprazole and DMP–777. Group 2A and 2B are dosed with DMP–777 once daily on Study Day 3 or Days 3 and 4,respectively, and receive control vehicle instead of omeprazole. Groups 3A and 3B are treated with omeprazole twice daily on Study Days 1 to 3 or Days 1 to 4, respectively, and receive control vehicle instead of DMP–777. Groups 4A and 4B are dosed with bothomeprazole and DMP–777. On Study Days 1 and 2, animals are pretreated with omeprazole twice daily, the dosing intervals separated by approximately 6 hr. On Study Day 3 (Group 4A) or Days 3 and 4 (Group 4B), omeprazole is coadministered with DMP–777. The first dose of omeprazole is administered approximately 1 hr prior to the dose of DMP–777. The second dose is approximately 6 hr after the last dose of DMP–777. Groups 1A, 2A, 3A, and 4A are sacrificed on Day 4. Groups 1B, 2B, 3B, and 4B are sacrificed on Day 5.Bromodeoxyuridine (BrdU) is administered by intraperitoneal injection to all the rats, 2 hr prior to necropsy.References:[1]. Ogawa M, et al. Omeprazole treatment ameliorates oxyntic atrophy induced by DMP–777. Dig Dis Sci. 2006 Mar;51(3):431–9.[2]. Zagrobelny J, et al. Separation of the four stereoisomers of a potent inhibitor (L–694,458) of human leukocyte elastase and its determination in human plasma using achiral/chiral chromatography with column switching. J Pharm Biomed Anal. 1998 Sep 1;17(6–7[3]. Weis VG, et al. Maturity and age influence chief cell ability to transdifferentiate into metaplasia. Am J Physiol Gastrointest Liver Physiol. 2016 Nov 23:ajpgi.00326.2016.Product Name:DMP 777Cat. No.:HY-75957CAS No.:157341-41-8Molecular Formula:C 31H 40N 4O 6Molecular Weight:564.67Target:Elastase Pathway:Metabolic Enzyme/Protease Solubility:10 mM in DMSOCaution: 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。
致癌染料检测DIN 54231 致癌染料测试DIN 54231Allergenous disperse dyes 致敏性分散染料 HPLC-MS , DIN 54231Carcinogen dyes 致癌染料 HPLC-MS , DIN 54231一、致敏性分散染料Allergenous disperse致敏染料是指某些会引起人体或动物的皮肤、黏膜或呼吸道过敏的染料。
人体吸入性的过敏主要集中于呼吸道和粘膜,部分活性染料(可分为颗粒状和液状)可造成此类致敏。
目前致敏染料共发现27种。
在国际知名生态安全规范Oeko-Tex Standard 100的2008版中,将其中的20种致敏性分散染料列为生态纺织品的监控项目,下表中1#~20#,并增加了另外两种致敏染料,21#和22#。
这些染料主要用于聚酯、聚酰胺和醋酯纤维的染色,其中的17种为早期用于醋酸纤维的分散染料。
欧盟于2002年5月推出的Eco-Label 标签标准规定:该标准所列出的17种染料(比Oeko-Tex Standard 100少了3种:C.I. 分散蓝1、C.I. 分散棕1和C.I. 分散黄3),并规定当染色纺织品的耐汗渍色牢度(酸性和碱性)低于4级时,不得使用。
了解其他相关及检测请进个人主页禁用的致敏染料编号染料英文名称染料中文名称CAS No.1# Disperse Blue 1 分散蓝1 2475-45-82# Disperse Blue 3 分散蓝3 2475-46-93# Disperse Blue 7 分散蓝7 3179-90-64# Disperse Blue 26 分散蓝26 3860-63-75# Disperse Blue 35 分散蓝35 12222-75-26# Disperse Blue 102 分散蓝102 69766-79-67# Disperse Blue 106 分散蓝106 12223-01-78# Disperse Blue 124 分散蓝124 61951-51-79# Disperse Brown 1 分散棕1 23355-64-810# Disperse Orange 1 分散橙 1 2581-69-311# Disperse Orange 3 分散橙3 730-40-512# Disperse Orange 37/76 分散橙37/76 13301-61-613# Disperse Red 1 分散红1 2872-52-814# Disperse Red 11 分散红11 2872-48-215# Disperse Red 17 分散红17 3179-89-316# Disperse Yellow 1 分散黄1 119-15-317# Disperse Yellow 3 分散黄3 2832-40-818# Disperse Yellow 9 分散黄 9 6373-73-519# Disperse Yellow 39 分散黄39 12236-29-220# Disperse Yellow 49 分散黄49 54824-37-221# Disperse Yellow 23 分散黄23 6250-23-322# Disperse Orange 149 分散橙149 85136-74- 9国际上目前对纺织品上限用致敏性分散染料检出的限定值,不同的买家所制订的合格性评定标准存在一些差异。
美国贝克曼库尔特流式细胞分析仪(Beckman coulter cell)产品型号:Cell Lab Quanta SC当前价格:0.00元产品数量:0新旧程度:全新有效期至:0000-00-00所在地:产品简介:仪器简介:T细胞亚群检测的CD45/CD4/CD8/CD3、CD45/CD56/CD19/CD3;阵发性血红蛋白尿(PNH)检测的CD55、CD59;血小板无力症(GT)检测的CD41、CD61等等详细信息仪器简介:T细胞亚群检测的CD45/CD4/CD8/CD3、CD45/CD56/CD19/CD3;阵发性血红蛋白尿(PNH)检测的CD55、CD59;血小板无力症(GT)检测的CD41、CD61等等。
但对于白血病/淋巴瘤免疫分型,国际上迄今为止也没有统一的抗体组合。
在2000年国际细胞分析学会(ISAC)大会上,临床血细胞计数协会组织了一次国际专家会议,以期对检测血液淋巴系统肿瘤所需最少、最有效的单抗数达成共识。
75%与会者一致认为,对于慢性淋巴系统增殖性疾病(CLD)有9种单抗:CD5,CD19,κ,λ,CD3,CD20,CD23,CD10,CD45对初诊来说是最基本的。
淋巴瘤和CLD相似,需要至少12-16种单抗。
对于急性白血病(AL),75%的与会者认为大约13-15种单抗是最基本的:CD10,CD19,CD79a,CD13,CD33,CD34,CD45,CD2,MPO,CD7,CD14,CD3,HLA-DR等,对初步鉴别白血病系列是必需的。
其他一些(CD16,CD56,CDw65,TdT,cyCD3)可能对某些病例有用。
几乎所有的投票者都认为,要对急性白血病完善分类所需单抗的恰当数量平均为20-24种。
但这些抗体之间组合也是一大难题,目前也无统一规定(如表二)。
大会多数发言者(11/13)指出,对已确诊病人的监护和分期来说,仅需较少单抗。
抗体的质量控制是实验的关键环节。
抗体的质量包括其特异性、灵敏度、精密度。