eppendorfepTIPS独立包装吸头,50-1000μl,生物纯级,100个_盒
京聚合美生物科技有限公司 M5 Endofree Pureplasmid Maxi Kit 使用说明
M5Endofree Pureplasmid Maxi Kit使用说明书产品名称单位货号M5Endofree Pureplasmid Maxi Kit10T MF032-01【储存条件】本试剂盒在室温储存12个月不影响使用效果。
内毒素清除剂可以常温运输，4˚C保存一个月，长期保存请放-20˚C。
【产品简介】本试剂盒采用改进SDS-碱裂解法裂解细胞，粗提物通过独特的内毒素清除剂选择性结合离心除去内毒素，然后离心吸附柱内的硅基质膜在高盐、低pH值状态下选择性地结合溶液中的质粒DNA，再通过去蛋白液和漂洗液将杂质和其它细菌成分去除，最后低盐、高pH值的洗脱缓冲液将纯净质粒DNA从硅基质膜上洗脱。
【产品特色】1.离心吸附柱内硅基质膜全部采用Whatman特制吸附膜，柱与柱之间吸附量差异极小，可重复性好。
2.不需要苯酚，氯仿等试剂，也不需要乙醇沉淀，快速方便。
从100-200ml大肠杆菌LB培养液中，可快速提取0.2-0.5mg纯净的高拷贝质粒DNA，提取率达80-90%。
3.独特工艺配方清除内毒素，内毒素含量极低（<0.1EU/μg DNA），细胞转染效果极佳。
也可直接用于酶切、转化、PCR、体外转录、测序等各种分子生物学实验。
【产品组份】10T注意事项细胞悬浮液（S1）77ml初次使用前请按瓶标说明加入所有的RNase A于S1中，4˚C保存细胞裂解液（S2）77ml用毕立即盖紧瓶盖；如有结晶析出，可于55˚C水浴加热助溶中和缓冲液（S3）77ml有刺激性，请勿直接接触皮肤浓缩漂洗液（WB）2x25ml初次使用前请按瓶标说明加入无水乙醇混匀洗脱缓冲液（EB）20mlRNase A溶液（10mg/ml）750μl-20˚C长期保存内毒素清除剂25ml-20˚C长期保存离心吸附柱DC10个室温密闭干燥保存50ml收集管10个室温密闭干燥保存【实验准备】用户需自行准备的材料：含适当抗生素的LB培养基，无水乙醇，台式离心机。
毕赤酵母表达知识归纳
毕赤酵母表达知识归纳1a.配制500×BIOTIN stock solution（0.02％）有这么3种方案：1、懒人是将Biotin直接溶在去离子水中，放过夜，基本就能溶；2、急性子是将溶液配成0.02N的NaOH，就很容易溶解了；3、水浴加热，温度不能高于50度。
D－生物素是具有生物活性的生物素，也就是vitaminH。
在毕赤酵母代谢过程中，作为多种酶的辅基起作用。
天然培养基中一般可以不单独添加，因为YNB中、酵母粉、蛋白胨中均含有一定量的生物素，但是做高密度发酵还是必须要添加的。
b.有几个比较迷惑的问题请教大家：(很典型的小问题）1、制感受态细胞，OD多少比较好？pyrimidine 战友的方法：取1mlGS115过夜培养物(OD约6-10) 分装到1.5ml EP管中。
说明书还有一些文献是说在1.3左右效率高，再高了效率会很低2、关于高效转化法，文献说用(LiAc)，而invitrogen的说明书说转化毕赤酵母用(LiAc)没用，要用LiCl。
Lithium acetate does not work with Pichia pastoris. Use only lithium chloride.3、YNB到底能高温灭么？有的说能有的说不能。
过滤灭菌的怎么操作？我是把滤器装好膜绑到瓶口用纱布盖上，报纸包上，瓶盖放烧杯里单灭。
然后把配好的溶液用注射器一点点推进去。
4、葡萄糖为什么在YPD里一起灭颜色很深，单灭则不会。
该115度还是121度灭？网上搜了下，都有人用！5、电转化参数用400欧还是200欧？有的用400，有的还专门说不是用400。
都是从园里看到的！电击参数：1.5KV，25uF，200欧姆（不是400）6、电转后，在MD平板上长的应该就是整合了目的基因的重组子了吧？如果不想筛高拷贝的，是否PCR验证一下即可？网友的回答：ynb最好不灭菌，我是0.22um过滤处理的。
invitrogen手册上可以灭菌的。
逆转录试剂盒( transcriptor first strand cdna synthesis kit ) 说明书
3
04 379 012 001
Transcriptor First Strand cDNA Synthesis Kit y Version 6.0
1. What this Product Does, continued
Vial/ Cap
Label
Store the kit at ؊15 to ؊25°C N Store Control RNA (vial 7 in
Cat. No 04 379 012 001) at —70°C or below.
y Version 6.0
Content version: September 2010
9 (7 for b,c) colorless
Water, PCR-grade
Content
a) Cat. No. 04 379 012 001 b) Cat. No. 04 896 866 001 c) Cat. No. 04 897 030 001
a) 1 vial, 1 ml b) 2 vials, each 1ml c) 3 vials, each 1 ml
L In Cat. No. 04 896 866 001 and Cat. No. 04 897 030 001 the control reagents (vial 7 and 8) are not included. Therefore, in these kits vial 7 is Water, PCR Grade.
Random Hexamer Primer
a) 1 vial, 100 l (600 M) b) 1 vial, 200 l (600 M) c) 2 vials, each 200 l (600 M)
API20E（20100）
20 100 / 20 160 API 20 E 只供体外诊断用肠杆菌和其它革兰氏阴性杆菌鉴定系统API 20 E是肠杆菌科和其它G-杆菌的标准鉴定系统包括23个标准化微型生化测试和鉴定资料库。
可鉴定菌谱名录请参考鉴定表。
原理API 20 E试验条是由20个含干燥底物的小管所组成。
这些测定管用细菌悬浮液接种。
培养一定时间后通过代谢作用产生颜色的变化或是加入试剂后变色而观察其结果。
根据说明表判读反应参照分析图索引和APILAB Plus软件得到鉴定结果。
试剂一个ref. 2010025测试试剂盒包括25条API 20 E试验条25个培养盒25张报告单1个封口夹1份API 20E操作说明书一个ref. 20160100测试试剂盒包括: 100条API 20 E试验条100个培养盒100张报告单1个封口来1份API 20 E说明书附加试剂不包括在试剂盒内NaCl 0.85 悬浮液 5 ml Ref. 20230或Suspension Medium县浮液5 ml Ref. 20150 API 20 E试剂盒Ref. 20120 TDAINDVP 1VP 2NIT 1NIT 2和OX或单一附加试剂: TDA Ref. 70400 JAMES Ref. 70540 VP 1 Ref. 70420 VP 2 Ref. 70430 NIT 1 Ref. 70440 NIT 2 Ref. 70450 OX Ref. 70460 Zn试剂Ref. 70380 石腊油Ref. 70100 API专用一次性加样管Ref. 70250 API LAB PLUS鉴定结果分析软件附加试剂安瓿架Ref. 70200 必要时需要以下附加试剂API OF Medium Ref. 50110 测定其发酵型或氧化型代谢作用API M Medium Ref. 51020 测定兼性厌氧细菌的运动性。
所需一般实验室设备培养箱3537°C 冰箱本生煤气喷灯或酒精灯记号笔培养基和试剂成份NaCl 0.85 培养基 5 ml or 悬浮基物5 ml NaCl 8.5 g 无离子水1000 ml 无离子水TDA 试剂 5 ml Fecl3 3.4 g H2O 加至100 ml JAMES 试剂5 ml J 2183化合物保密0.5 g HCl 1N 加至100 ml S24/25: 避免接触皮肤和眼.如与眼接触必须立即用水冲洗至少10分钟。
ELISA
酶联免疫吸附试验（ELISA）作者：发布日期：(2009-05-31) 浏览次数：7次酶联免疫吸附试验（enzyme linked immunosorbent assay，ELISA）是一种固相酶免疫测定技术。
它的基础是抗原或抗体的固相化及抗原或抗体的酶标记。
结合在固相载体表面的抗原或抗体仍保持其免疫学活性，酶标记的抗原或抗体既保留其免疫学活性，又保留酶的活性。
在测定时，受检标本（测定其中的抗体或抗原）与固相载体表面的抗原或抗体起反应。
用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。
再加入酶标记的抗原或抗体，也通过反应而结合在固相载体上。
此时固相上的酶量与标本中受检物质的量呈一定的比例。
加入酶反应的底物后，底物被酶催化成为有色产物，产物的量与标本中受检物质的量直接相关，故可根据呈色的深浅进行定性或定量分析。
ELISA的主要类型有双抗体夹心法、间接法、竞争法、捕获法等。
试剂准备1．包被液（碳酸盐缓冲液pH9.6）：Na2CO3 1.59g；NaHCO3 2.93g；加800ml水溶解，定容至1000ml加0.02%NaN3，0.22μm膜过滤，4℃保存。
2．洗涤缓冲液（PBST）：NaCl 9g；KCl 0.25g；Na2HPO4 3.63g；KH2PO4 0.25g；加水溶解，定容至1000ml加0.2% Tween20混匀。
3．封闭液：10%小牛血清（1ml小牛血清加到9ml 1×PBS中混匀）。
4．底物缓冲液（磷酸盐-柠檬酸缓冲液pH6.0）：Na2HPO4 56.77g；柠檬酸5.6g；加水800ml溶解，定容至1000ml，过滤0.22μm膜，4℃保存。
5．终止液：1mol/L H2SO4。
一、双抗体夹心法检测抗原双抗体夹心法用于检测抗原。
它是利用待测抗原上的两个抗原决定簇A和B分别与固相载体上的抗体A和酶标记抗体B结合，形成抗体A-待测抗原-酶标抗体B复合物，复合物的形成量与待测抗原含量成正比。
苏州科铭试剂盒总清单-5.3
咨询 800 420 2300 1200 1000 520 1100 580 1550 800 1200 620
L-半乳糖苷-1,4-内酯脱氢酶(Gal LDH)测试盒
L-半乳糖苷-1,4-内酯脱氢酶(Gal LDH)测试盒
抗坏血酸氧化酶（AAO）测试盒
抗坏血酸氧化酶（AAO）测试盒
抗坏血酸过氧化物酶（APX）测试盒
抗坏血酸过氧化物酶（APX）测试盒
单脱氢抗坏血酸还原酶（MDHAR）测试盒
单脱氢抗坏血酸还原酶（MDHAR）测试盒
微量法
紫外分光光度法
微量法
100管/48样 50管/24样 50管/48样 100管/96样 50管/48样 100管/96样 50管/48样 100管/96样 50管/48样 100管/96样 50管/48样 100管/96样 50管/48样 100管/96样 50管/48样
100管/96样
50管/48样
丙酮酸脱羧酶（PDC）测试盒
丙酮酸脱羧酶（PDC）测试盒
醇脱氢酶（ADH）测试盒
醇脱氢酶（ADH）测试盒
单脱氢抗坏血酸还原酶（MDHAR）测试盒
单脱氢抗坏血酸还原酶（MDHAR）测试盒
乙醛脱氢酶（ALDH）测试盒 乙醛脱氢酶（ALDH）测试盒
烟酸含量测试盒
微量法 可见分光光度法 高效液相色谱法
微量法 可见分光光度法
脱氢抗坏血酸还原酶（DHAR）测试盒
脱氢抗坏血酸还原酶（DHAR）测试盒
肝素钠_usp33
Record the H NMR spectra of the Standard solution and System suitability solution at 25 . 1 Collect the H NMR spectrum with a spectral window of at least 10 to 2 ppm and without spinning. The number of transients should be adjusted until the signal-to-noise ratio of the Nacetyl heparin signal in the Standard solution is at least 1000/1 in the region near 2 ppm. The Standard solution shall be run at least daily when Sample solutions are being run. For all samples, the TSP methyl signal should be set to 0.00 ppm. The chemical shift for the Nacetyl resonance of heparin and oversulfated chondroitin sulfate in the System suitability solution should be observed at 2.05 ± 0.02 and 2.16 ± 0.03 ppm, respectively. Record the H NMR spectrum of the Sample solution at 25 . Draw a baseline from 8.00 ppm to 0.10 ppm. The ppm values for H1 of GlcNAc/GlcNS, 6S (signal 1), H1 of IdoA2S (signal 2), the H2 of GlcNS (signal 3), and the methyl of GlcNAc (signal 4) of heparin are present at 5.42, 5.21, 3.28 (doublet centered at 3.28 ppm), and 2.05 ppm, respectively.1 The ppm values of these signals do not differ by more than ±0.03 ppm. Measure the signal heights above the baseline of signal 1 and signal 2, and calculate the mean of these signal heights. Other signals of variable heights and ppm values, attributable to heparin and HOD, may be seen between signal 2 and 4.55 ppm. Residual solvent signals may be observed in the 0.10–3.00 range. Heparin Sodium must meet the requirements stated in Residual Solvents 467 . Suitability requirements Number of transients: Adjust until the signal-to-noise ratio of the N-acetyl heparin signal in the Standard solution is at least 1000/1 in the region near 2 ppm. Chemical shift: The TSP methyl signal should be set to 0.00 ppm for all samples. Chemical shifts (for the N-acetyl resonance of heparin and oversulfated chondroitin sulfate): Should be observed at 2.05 ± 0.02 and 2.16 ± 0.03 ppm, respectively, System suitability solution Analysis Sample: Sample solution Acceptance criteria: No unidentified signals greater than 4% of the mean of signal height of 1 and 2 are present in the following ranges: 0.10–2.00, 2.10–3.20, and 5.70–8.00 ppm. No signals greater than 200% signal height of the mean of the signal height of 1 and 2 are present in the 3.35–4.55 ppm for porcine heparin . • B. CHROMATOGRAPHIC IDENTITY Solution A: Dissolve 0.8 g of monobasic sodium phosphate dihydrate in 2 L of water and adjust with phosphoric acid to a pH of 3.0. Pass the solution through a filter membrane with pore sizes of 0.45 µm and degas before use. Solution B: Dissolve 0.8 g of monobasic sodium phosphate dihydrate and 280 g of sodium perchlorate monohydrate in 2 L of water and adjust with phosphoric acid to a pH of 3.0. Pass the solution through a filter membrane with pore sizes of 0.45 µm and degas before use. Mobile phase: See the gradient table below. Time (min) 0 60 61 Solution A (%) 80 10 80 Solution B (%) 20 90 20 Elution Equilibration Linear gradient Linear gradient
(整理)什么是PCR.
聚合酶链反应（Polymerase Chain Reaction, PCR），是一种对特定的DNA片段在体外进行快速扩增的方法，也是分子生物学实验最常用到的一种方法。
根据适用对象的不同，又有荧光实时PCR、逆转录PCR等区别。
本专题详细介绍了PCR实验的基本原理、步骤及常见问题的解决办法。
第一部分:PCR的基础知识第一章:聚合酶链式反应(PCR)的历史和发展本文章内容包括:聚合酶链反应的历史回顾、聚合酶链反应相关技术的发展、其它体外核酸扩增技术、PCR技术的应用举例。
第一节：聚合酶链反应的历史回顾1、核酸体外扩增最早的设想由Khorana及其同事于1971年提出：“经过DNA变性，与合适引物杂交，用DNA聚合酶延伸引物，并不断重视该过程便可克隆tRNA基因”。
但由于当时很难进行测序和合成寡核苷酸引物，且当时(1970年)Smith等发现了DNA限制性内切酶，使体外克隆基因成为可能，所以，使Khorana等的早期设想被人们遗忘。
2、聚合酶链反应的发明直到1985年，美国PE-Cetus公司的人类遗传研究室Mullis等人才发明了具有划时代意义的聚合酶链反应(Polymerase Chain Reaction, PCR), 使人们梦寐以求的体外无限扩增核酸片段的愿望成为现实。
其原理类似于DNA的体内复制，只是在试管中给DNA的体外合成提供一种合适条件。
开始是使用大肠杆菌DNA聚合酶Klenow片段来扩增人基因组中的特异片段。
由于该酶不耐热，因此，每次加热变性DNA后都要重新补加Klenow酶。
在操作多份标本时，这一过程耗时，费力，且易出错。
耐热DNA聚合酶的应用使得PCR反应更易于自动化，继而PE-Cetus公司推出了第一台PCR热循环仪，使该技术的自动化成为现实。
Mullis等因此项技术于1993年获得诺贝尔奖金。
二、聚合酶链反应相关技术的发展PCR及其相关技术的发展速度是惊人的。
国际上分别于1988年和1990年在美国和英国召开了第一届和第二届PCR技术专题研讨会。
axykit 报价
Plasmid DNA purification AxyPrep-96 Plasmid Kit AxyPrep-96 质粒DNA试剂盒 1×96 prep Plasmid DNA purification AxyPrep-96 Plasmid Kit AxyPrep-96 质粒DNA试剂盒 24×96 prep Plasmid DNA purification AxyPrep-96 Plasmid Kit AxyPrep-96 质粒DNA试剂盒 4×96 prep PCR clean-up PCR clean-up PCR clean-up AxyPrep-96 PCR Clean-up AxyPrep-96 Kit PCR 1×96 清洁试剂盒 prep AxyPrep-96 PCR Clean-up AxyPrep-96 Kit PCR 24×96 清洁试剂盒 prep AxyPrep-96 PCR Clean-up AxyPrep-96 Kit PCR 4×96 清洁试剂盒 prep
Plasmid DNA purification AxyPrep Midi PlasmidAxyPrep Kits 质粒中量制备试剂盒 10 prep Plasmid DNA purification AxyPrep Midi PlasmidAxyPrep Kits 质粒中量制备试剂盒 25 prep Plasmid DNA purification AxyPrep Midi PlasmidAxyPrep Kits 质粒中量制备试剂盒 2 prep
AP-MN-BF-VNA-4G AP-MN-BF-VNA-50G AP-MN-BF-VNA-VN AP-MN-BL-GDNA-250G AP-MN-BL-GDNA-4G AP-MN-BL-GDNA-50G AP-MN-BL-GDNA-AP1 AP-MN-BL-GDNA-AP2 AP-MN-BL-GDNA-TE AP-MN-BL-RNA-250G AP-MN-BL-RNA-4G AP-MN-BL-RNA-50G AP-MN-BT-GDNA-250G AP-MN-BT-GDNA-250T AP-MN-BT-GDNA-4G AP-MN-BT-GDNA-50G AP-MN-EP-150G AP-MN-EP-4G AP-MN-EP-50G AP-MN-FEP-150G AP-MN-FEP-4G AP-MN-FEP-50G AP-MN-MiRNA-250G AP-MN-MiRNA-4G AP-MN-MiRNA-50G AP-MN-MS-GDNA-250G AP-MN-MS-GDNA-4G AP-MN-MS-GDNA-50G AP-MN-MS-RNA-250G AP-MN-MS-RNA-4G AP-MN-MS-RNA-50G AP-MN-P-250G AP-MN-P-250G-RA AP-MN-P-4G AP-MN-P-500G AP-MN-P-500-S1 AP-MN-P-500-S2 AP-MN-P-500-S3 AP-MN-P-500-W2 AP-MN-P-50G AP-MX-BL-GDNA-10G AP-MX-BL-GDNA-25G AP-MX-BL-GDNA-2G AP-MX-EP-10G AP-MX-EP-25G AP-MX-EP-2G AP-MX-MS-RNA-10G AP-MX-MS-RNA-25G
依诺肝素钠标准(USP 39)
•L IMIT OF F LUORIDE ADDITIONAL REQUIREMENTS [N OTE—Use plasticware throughout this test.]•P ACKAGING AND S TORAGE: Preserve in tight, light-resistant Buffer: Dissolve 110g of sodium chloride and 1g of containers, and avoid exposure to excessive heat.sodium citrate in 700mL of water in a 2000-mL volu-•USP R EFERENCE S TANDARDS〈11〉metric flask. Cautiously add 150g of sodium hydroxide,USP Enflurane RSand dissolve with shaking. Cool to room temperature,and, while stirring, cautiously add 450mL of glacialacetic acid to the cooled solution. Cool, add 600mL ofisopropyl alcohol, and dilute with water to volume; thepH of this solution is 5.0–5.5.Enoxaparin SodiumStandard stock solution: 1mg/mL of fluoride ion, pre-pared as follows. Transfer 221mg of sodium fluoride, previously dried at 150° for 4 h, to a 100-mL volumet-ric flask. Add 20mL of water, and mix to dissolve. Add 1.0mL of sodium hydroxide solution (1 in 2500), and dilute with water to volume. [N OTE—Store in a tightly closed plastic container.]Standard solution A: 100mL of a solution containing 1µg/mL of fluoride ion in Buffer, from Standard stock solutionStandard solution B: 100mL of a solution containing 3µg/mL of fluoride ion in Buffer, from Standard stock solutionStandard solution C: 100mL of a solution containing 5µg/mL of fluoride ion in Buffer, from Standard stock solutionStandard solution D: 100mL of a solution containing 10µg/mL of fluoride ion in Buffer, fromStandard stocksolution[9041-08-1].Sample stock solution: Enflurane and water (1:1).Shake the Sample stock solution for 5 min, allow theDEFINITIONliquids to separate completely, and use the water layer.Enoxaparin Sodium is the sodium salt of a depolymerized Sample solution: Sample stock solution and Bufferheparin. It is obtained by alkaline depolymerization of (1:1). Use volumetric glassware.heparin benzyl ester. The starting material, heparin, is ob-Electrode system: Use a pH meter capable of a mini-tained exclusively from porcine intestinal mucosa. Heparin mum reproducibility of ±0.2 mV, equipped with a glass-source material used in the manufacture of Enoxaparin sleeved, calomel-fluoride, specific-ion electrode systemSodium complies with the compendial requirements (see pH 〈791〉).stated in the Heparin Sodium monograph. Enoxaparin So-Analysisdium consists of a complex set of oligosaccharides that Samples: Standard solutions and Sample solutionhave not yet been completely characterized. The majority Transfer the solution to a 150-mL beaker, add aof the components have a 4-enopyranose uronate struc-polytef-coated stirring bar, and immerse the elec-ture at the nonreducing end of their chain. About 20% of trodes in the solution. Stir with a magnetic stirrerthe materials contain a 1,6-anhydro derivative on the re-having an insulated top until equilibrium is obtainedducing end of the chain, the range being between 15% (1–2 min), and record the potential, in mv. [N OTE—and 25%. The weight-average molecular weight of Enox-Rinse and dry the electrodes between measurements,aparin Sodium is 4500 Da, the range being between taking care to avoid damaging the crystal of the spe-3800 and 5000 Da; about 16% have a molecular weight cific-ion electrode.]of less than 2000 Da, the range being between 12.0% Plot the logarithm of the fluoride-ion concentrations,and 20.0%; about 74% have a molecular weight between in µg/mL, of the Standard solutions versus the poten-2000 and 8000 Da, the range being between 68.0% and tial, in mV. From the measured potential of the Sam-82.0%. NMT 18.0% have a molecular weight higher thanple solution and the standard curve, determine the8000 Da. When prepared as a solution, the solution is concentration, in µg/mL, of fluoride ions in the Sam-analyzed for clarity and degree of color using a validated ple solution.method. The degree of sulfation is NLT 1.8per disaccha-Acceptance criteria: NMT 10µg/mLride unit. It has a potency of NLT 90 and NMT 125 Anti-•L IMIT OF N ONVOLATILE R ESIDUEFactor Xa International Units (IU)/mg, and NLT 20.0 and Analysis: Allow 10.0mL to evaporate at room tempera-NMT 35.0 Anti-Factor IIa IU/mg, calculated on the dried ture in a tared evaporating dish, dry the residue at 50°basis. The ratio of Anti-Factor Xa activity to Anti-Factor IIa for 2 h, and weigh.activity is between 3.3 and 5.3.Acceptance criteria: NMT 2mg of residue remains.IDENTIFICATIONSPECIFIC TESTS•A. U LTRAVIOLET A BSORPTION〈197U〉•S PECIFIC G RAVITY〈841〉: 1.516–1.519Medium: 0.01 N hydrochloric acid•R EFRACTIVE I NDEX〈831〉: 1.3020–1.3038 at 20°Sample solution: 500µg/mL•W ATER D ETERMINATION, Method I〈921〉: NMT 0.14%Acceptance criteria: The spectra exhibit maxima at •A CIDITY OR A LKALINITY231±2 nm.Sample: Shake 20mL of Enflurane with 20mL of car-bon dioxide-free water for 3 min, and allow the layersto separate. Draw off the water layer, add bromocresol Change to read:purple TS as the indicator, and titrate with 0.010 Nsodium hydroxide or 0.010 N hydrochloric acid.•B.13C NMR S PECTRUMAcceptance criteria: NMT 0.10mL of 0.010 N sodium(See •Nuclear Magnetic Resonance Spectroscopy 〈761〉•(CN hydroxide or NMT 0.60mL of 0.010 N hydrochloric1-May-2016).)acid is required for neutralization.Standard solution: Dissolve 200mg of USP EnoxaparinSodium RS in a mixture of 0.2mL of deuterium oxideand 0.8mL of water. Add 0.05mL of deuterated meth-Acceptance criteria:M2000 is between 12.0% and anol to serve as an internal reference.20.0%, M2000–8000 is between 68.0% and 82.0%, and Sample solution: Dissolve 200mg of Enoxaparin So-M8000 is NMT 18.0%.dium in a mixture of 0.2mL of deuterium oxide and•E. I DENTIFICATION T ESTS—G ENERAL, Sodium〈191〉: Meets0.8mL of water. Add 0.05mL of deuterated methanol.the requirementsAnalysis: Transfer the Standard solution and the SampleASSAYsolution to NMR tubes of 5-mm diameter. Using apulsed (Fourier transform) NMR spectrometer operatingat NLT 75MHz for 13C, record the 13C NMR spectra of Change to read:the Standard solution and the Sample solution at 40°.Acceptance criteria: The spectra are similar.•ANTI-F ACTOR X a A CTIVITY•C. The ratio of the numerical value of the Anti-Factor Xa Acetic acid solution: Glacial acetic acid and water activity, in Anti-Factor Xa IU/mg, to the numerical value(42:58)of the Anti-Factor IIa activity, in Anti-Factor IIa IU/mg, as pH 7.4polyethylene glycol 6000 buffer: Dissolve determined by the Assay (Anti-Factor Xa Activity) and Im- 6.08g of tris(hydroxymethyl)aminomethane and 8.77g purities (Anti-Factor IIa Activity), respectively, is NLT 3.3of sodium chloride in 500mL of water. Add 1.0g of and NMT 5.3.polyethylene glycol 6000, adjust with hydrochloric acid •D. M OLECULAR W EIGHT D ISTRIBUTION AND W EIGHT-A VERAGE to a pH of 7.4, and dilute with water to 1000mL.M OLECULAR W EIGHT pH 7.4 buffer: Dissolve 6.08g of tris(hydroxymethyl)-Mobile phase: Prepare a 0.5 M lithium nitrate solution.aminomethane and 8.77g of sodium chloride in Pass through a membrane filter of 0.45-µm or smaller500mL of water. Adjust with hydrochloric acid to a pH pore size, and degas with helium.of 7.4, and dilute with water to 1000mL.Standard solution: 10mg/mL of USP Enoxaparin So-pH 8.4 buffer: Dissolve 3.03g of tris(hydroxymethyl)-dium RS in Mobile phase aminomethane, 5.12g of sodium chloride, and 1.40g Sample solution: 10mg/mL of Enoxaparin Sodium in of edetate sodium in 250mL of water. Adjust with hy-Mobile phase drochloric acid to a pH of 8.4, and dilute with water to Chromatographic system500mL.(See Chromatography 〈621〉, System Suitability.)Human antithrombin III solution: Reconstitute a vial Mode: Size exclusion LC of antithrombin III (see Reagents, Indicators, and Solu-Detector: Differential refractive index tions—Reagent Specifications) in water to obtain a solu-Column tion containing 5 Antithrombin III Units/mL. Dilute this Guard: 6-mm × 40-mm; packing L59solution with pH 7.4polyethylene glycol 6000 buffer to Analytical: Two 7.8-mm × 300-mm columns in series;obtain a solution having a concentration of 1.0 Anti-packing L59thrombin III Unit/mL.Temperature: Room temperature Factor Xa solution: Reconstitute a weighed quantity of Flow rate: 0.6mL/min maintained constant to ± 1.0%bovine factor Xa (see Reagents, Indicators, and Solu-Analysis: Reconstitute 1 vial each of USP Enoxaparin tions—Reagent Specifications) in pH 7.4polyethylene gly-Sodium Molecular Weight Calibrant A RS and USP col 6000 buffer to obtain a solution that gives an in-Enoxaparin Sodium Molecular Weight Calibrant B RS in crease in absorbance value at 405 nm of NMT 0.20 1mL of Mobile phase. Separately inject 20µL of USP absorbance units/min when assayed as described below Enoxaparin Sodium Molecular Weight Calibrant A RS but using as an appropriate volume, V, the volume in and USP Enoxaparin Sodium Molecular WeightµL of pH 7.4 buffer instead of VµL of the enoxaparin Calibrant B RS, record the chromatograms, and meas-solution.ure the retention times. Inject in duplicate 20µL each Chromogenic substrate solution: Prepare a solution of of the Standard solution and the Sample solution, and a suitable chromogenic substrate for amidolytic test record the chromatograms for a length of time to en-(see Reagents, Indicators, and Solutions—Reagent Specifi-sure complete elution, including salt and solvent peaks.cations) for factor Xa in water to obtain a concentration Calculate the total peak areas under each of the Stan-of about 3 mM. Dilute with pH 8.4 buffer to obtain a dard solution and Sample solution chromatograms, ex-solution having a concentration of 0.5 mM.cluding salt and solvent peaks at the end.Standard solutions: Reconstitute the entire contents of Calibration curve: Plot the retention times on the x-an ampul of USP Enoxaparin Sodium for Bioassays RS axis against the peak molecular weights on the y-axis with water, and dilute with pH 7.4 buffer to obtain four for the peaks from USP Enoxaparin Sodium Molecular dilutions in the concentration range between 0.025 and Weight Calibrant A RS and USP Enoxaparin Sodium Mo-0.2 Anti-Factor Xa IU/mL.lecular Weight Calibrant B RS, and fit the data to a Sample solutions: Proceed as directed for Standard so-third-order polynomial, using suitable gel permeation lutions to obtain concentrations of Enoxaparin Sodium chromatography (GPC) software.similar to those obtained for the Standard solutions.Calculations: Compute the data, using the same GPC Analysissoftware; determine the weight-average molecular Samples:Standard solutions, Sample solutions, Human weight, M w, for each of the duplicate chromatograms of antithrombin III solution, pH 7.4 buffer, Factor Xa solu-the Standard solution and the Sample solution; and take tion, Chromogenic substrate solution, and Acetic acid the average for each solution. Correct the mean value solutionof M w to the nearest 50. The Chromatographic system is Label 18 suitable tubes: B1 and B2 for blanks; T1, T2, suitable if M w for USP Enoxaparin Sodium RS is within T3, and T4 each in duplicate for the dilutions of the 150 Da of the labeled M w value. The M w for the Sample Sample solutions; and S1, S2, S3, and S4 each in dupli-solution is between 3800 and 5000 Da. Using the same cate for the dilutions of the Standard solutions. [NOTE—software, determine for each of the duplicate Sample Treat the tubes in the order B1, S1, S2, S3, S4, T1, T2,solution chromatograms the percentage of Enoxaparin T3, T4, T1, T2, T3, T4, S1, S2, S3, S4, B2.] To each Sodium chains with molecular weights lower than 2000tube add the same volume, V (20–50µL), of Human Da, M2000, the percentage of Enoxaparin Sodium chains antithrombin III solution and an equal volume, V, of with molecular weights in the range 2000–8000 Da,either the blank (pH 7.4 buffer) or an appropriate dilu-M2000–8000, and the percentage of Enoxaparin Sodium tion of the Sample solutions or the Standard solutions.chains with molecular weights greater than 8000 Da,Mix, but do not allow bubbles to form. Incubate at M8000. Average the duplicate values, and express to the37° for 1.0 min. Add to each tube 2V (40–100µL) of nearest 0.5%.Factor Xa solution, and incubate for 1.0 min. Add a 5V Analysis(100–250µL) volume of Chromogenic substrate solu-Samples:Standard solution A, Standard solution B, tion. Stop the reaction after 4.0 min with a 5V Standard solution C, Cesium chloride solution, and Sam-(100–250µL) volume of Acetic acid solution. Measure ple solutionthe absorbance of each solution at 405 nm, using a Concomitantly determine the absorbances of the Ce-suitable spectrophotometer (see •Ultraviolet-Visible sium chloride solution (blank), Sample solution, and Spectroscopy 〈857〉•(CN 1-May-2016)) against blank B1. The Standard solutions at 330.3 nm, using a sodiumreading of blank B2 relative to blank B1 is NMT ± 0.05hollow-cathode lamp and an air–acetylene flame. Us-absorbance unit.ing the absorbances of Standard solutions A–C, deter-Calculations: For each series, calculate the regression mine the sodium content in the Sample solution after of the absorbance against log concentrations of the an appropriate blank correction.Sample solutions and of the Standard solutions, and cal-Acceptance criteria: 11.3%–13.5% on the dried basis culate the potency of the Enoxaparin Sodium in IU ofIMPURITIESAnti-Factor Xa activity/mL, using statistical methods forparallel-line assays. The four independent log relativepotency estimates are then combined to obtain the Delete the following:final geometric mean. Its confidence limits are calcu-lated. Express the Anti-Factor Xa activity of Enoxaparin••H EAVY M ETALS, Method I〈231〉: NMT 30µg/g, using a Sodium/mg. 2.7% solution in water•(Official 1-Jan-2018) Acceptance criteria: The potency is NLT 90 and NMT125 Anti-Factor Xa IU/mg on the dried basis.SPECIFIC TESTS•P H 〈791〉: 6.2–7.7 for a 10.0% solution in water OTHER COMPONENTS•LOSS ON D RYING〈731〉: Dry 1g in a vacuum at 70° for 6•B ENZYL A LCOHOL C ONTENT h: it loses NMT 10.0% of its weight.Mobile phase: Acetonitrile, methanol, and water(3:1:16)Standard solution: 0.1mg/mL of USP Benzyl Alcohol Change to read:RS in waterSample solution: Weigh 0.5g of Enoxaparin Sodium•S PECIFIC A BSORBANCEinto a 10-mL volumetric flask, and dissolve in 5.0mL of(See •Ultraviolet-Visible Spectroscopy 〈857〉•(CN 1-May-2016).) 1N sodium hydroxide. Allow to stand at room temper-Sample solution: 0.5mg/mL of Enoxaparin Sodium in ature for about 1 h. Add 1.0mL of glacial acetic acid,0.01 N hydrochloric aciddilute with water to volume, and mix.Analysis: Obtain the UV spectra of the Standard solution Chromatographic system and the Sample solution between 200 nm and 300 nm (See Chromatography 〈621〉, System Suitability.)against a 0.01 N hydrochloric acid blank.Mode: LC Calculate the specific absorbance at the wavelength of Detector: UV 256 nm maximum absorbance at 231±2 nm, with reference Column: 4.6-mm × 15-cm stainless steel; packing L7to the dried substance:Flow rate: 1.0mL/min, maintained constant to ±10%Injection volume: 20µL Result = A× 100×1000/[M×l× (100 −E)] AnalysisA= absorbance at the wavelength of maximum Samples:Standard solution and Sample solutionabsorbanceCalculate the percentage of benzyl alcohol in the por-M= weight of Enoxaparin Sodium in the Sample tion of Enoxaparin Sodium taken:solution (mg)Result = (r U/r S) × (C S/C U) × 100l= path length (typically 1cm)E= loss on drying (%)r U= peak area of benzyl alcohol from the Sample Acceptance criteria: 14.0–20.0 on the dried basis solution•B ACTERIAL E NDOTOXINS T EST〈85〉: It contains NMT 0.01 r S= peak area of benzyl alcohol from the Standard USP Endotoxin Unit/IU of Anti-Factor Xa activity.solution•A NTI-F ACTOR II a A CTIVITYC S= concentration of benzyl alcohol in the Acetic acid solution, pH 7.4polyethylene glycol 6000Standard solution (mg/mL)buffer, pH 7.4 buffer, pH 8.4 buffer, and HumanC U= concentration of Enoxaparin Sodium in the antithrombin III solution: Proceed as directed in theSample solution (mg/mL)Assay for Anti-Factor Xa Activity, except that the concen-Acceptance criteria: NMT 0.1%tration of the Human antithrombin III solution is 0.5•N ITROGEN D ETERMINATION, Method II〈461〉: 1.8%–2.5%Antithrombin III Unit/mL.on the dried basis Thrombin human solution: Reconstitute thrombinhuman (see Reagents, Indicators and Solutions—ReagentSpecifications) in water, and dilute in pH 7.4polyethylene Change to read:glycol 6000 buffer to obtain a solution having a concen-tration of 5 Thrombin Units/mL.•S ODIUM C ONTENT Chromogenic substrate solution: Prepare a solution of (See •Atomic Absorption Spectroscopy 〈852〉•(CN 1-May-2016).) a suitable chromogenic substrate for an amidolytic test Cesium chloride solution: 1.27mg/mL of cesium chlo-(see Reagents, Indicators, and Solutions—Reagent Specifi-ride in 0.1 N hydrochloric acid cations) for thrombin in water to obtain a concentration Standard solution A: 0.0025% of sodium chloride in of about 3 mM. Immediately before use, dilute with pH Cesium chloride solution8.4 buffer to 0.5 mM.Standard solution B: 0.0050% of sodium chloride in Standard solutions: Reconstitute the entire contents of Cesium chloride solution an ampul of USP Enoxaparin Sodium for Bioassays RS Standard solution C: 0.0075% of sodium chloride in with water, and dilute with pH 7.4 buffer to obtain four Cesium chloride solution dilutions having concentrations in the range between Sample solution: Transfer 50.0mg of Enoxaparin So-0.015 and 0.075 IU of Anti-Factor IIa activity/mL.dium to a 100-mL volumetric flask, and dissolve in anddilute with Cesium chloride solution to volume.Sample solutions: Proceed as directed under Standard ADDITIONAL REQUIREMENTSsolutions to obtain concentrations of Enoxaparin Sodium•P ACKAGING AND S TORAGE: Preserve in tight containers, similar to those obtained for the Standard solutions.and store below 40°, preferably at room temperature.Analysis: Proceed as directed in the Assay for Anti-Factor•USP R EFERENCE S TANDARDS〈11〉Xa Activity, except to use Thrombin human solution in-USP Benzyl Alcohol RSstead of Factor Xa solution and to use Human antithrom-USP Endotoxin RSbin III solution as described P Enoxaparin Sodium RSCalculations: For each series, calculate the regression of USP Enoxaparin Sodium Molecular Weight Calibrant A RS the absorbance against log concentrations of the Sam-USP Enoxaparin Sodium Molecular Weight Calibrant B RS ple solutions and of the Standard solutions, and calculate USP Enoxaparin Sodium for Bioassays RSthe potency of the Enoxaparin Sodium in IU of Anti-Factor IIa activity/mg, using statistical methods for par-allel-line assays. The four independent dilution estimatesare then combined to obtain the final weighted mean.Then calculate the confidence limits. Express the Anti-Enoxaparin Sodium InjectionFactor IIa activity of Enoxaparin Sodium/mg.Acceptance criteria: It has a potency of NLT 20.0 and DEFINITIONNMT 35.0 Anti-Factor IIa IU/mg on the dried basis.Enoxaparin Sodium Injection is a sterile solution of Enox-•M OLAR R ATIO OF S ULFATE TO C ARBOXYLATE aparin Sodium in Water for Injection. Its appearance is Mobile phase: Carbon dioxide-free water analyzed for clarity and degree of color, using a validated Sample solution: 5mg/mL of Enoxaparin Sodium in method. Its potency value is NLT 90% and NMT 110% of carbon dioxide-free water the potency stated on the label in terms of International Chromatographic system Anti-Factor Xa Units (IU). It may contain, in multiple-dose (See Chromatography 〈621〉, System Suitability.)containers, a suitable antimicrobial preservative, such as Mode: LC benzyl alcohol.Detector: IonColumn: Two columns; one 1.5-cm × 2.5-cm column,IDENTIFICATIONpacked with an anion-exchange resin L64packing; and•A.one 1.5-cm × 7.5-cm column, packed with a cation-Analysis: Transfer the total contents of a single-dose exchange resin L65packing. The outlet of the anion-container or 0.4mL from a multiple-dose container to a exchange column is connected to the inlet of the cat-glass test tube, add 2mL of water and 1mL of 2% ion-exchange column.(w/v) protamine sulfate solution, and mix.Flow rate: 1mL/min Acceptance criteria: A creamy white precipitate is Analysis formed.Sample:Sample solution•B. U LTRAVIOLET A BSORPTION〈197U〉[N OTE—Regenerate the anion-exchange column and the Medium: 0.01 N hydrochloric acidcation-exchange column with 1N sodium hydroxide Standard solution: 500µg/mLand 1N hydrochloric acid, respectively, between two Sample solution: Transfer the total content of a single-injections.]dose container or 0.4mL from a multiple-dose con-Inject the Sample solution into the anion-exchange col-tainer to a 100-mL volumetric flask. Dilute with Medium umn, and collect the eluate from the cation-exchange to volume.column in a beaker at the outlet until the ion detector Acceptance criteria: The spectra exhibit maxima at reading returns to the baseline value. Quantitatively231±2 nm.transfer the eluate to a titration vessel containing a•C. I DENTIFICATION T ESTS—G ENERAL, Sodium〈191〉: Meets magnetic stirring bar, and dilute with carbon dioxide-the requirementsfree water to about 60mL. Position the titration vesselASSAYon a magnetic stirrer, and immerse the electrodes.Note the initial conductivity reading, and titrate withapproximately 0.1 N sodium hydroxide added in Change to read:100-µL portions. [N OTE—Prepare the sodium hydroxidesolution in carbon dioxide-free water.] Record the bu-•A NTI-F ACTOR X a A CTIVITYret reading and the conductivity meter reading after Acetic acid solution: Glacial acetic acid and water each addition of the sodium hydroxide solution.(42:58)Calculations: Plot the conductivity measurements on pH 7.4polyethylene glycol 6000 buffer: Dissolve the y-axis against the volumes of sodium hydroxide 6.08g of tris(hydroxymethyl)aminomethane and 8.77g added on the x-axis. The graph will have three linear of sodium chloride in 500mL of water. Add 1.0g of sections—an initial downward slope, a middle slight polyethylene glycol 6000, adjust with hydrochloric acid rise, and a final rise. For each of these sections draw the to a pH of 7.4, and dilute with water to 1000mL.best-fit straight lines, using linear regression analysis. At pH 7.4 buffer: Dissolve 6.08g of tris(hydroxymethyl)-the points where the first and second straight lines in-aminomethane and 8.77g of sodium chloride in tersect and where the second and third lines intersect,500mL of water. Adjust with hydrochloric acid to a pH draw perpendiculars to the x-axis to determine the of 7.4, and dilute with water to 1000mL.volumes of sodium hydroxide taken up by the sample pH 8.4 buffer: Dissolve 3.03g of tris(hydroxymethyl)-at those points. The point where the first and second aminomethane, 5.12g of sodium chloride, and 1.40g lines intersect corresponds to the volume of sodium hy-of edetate sodium in 250mL of water. Adjust with hy-droxide taken up by the sulfate groups (V S). The point drochloric acid to a pH of 8.4, and dilute with water to where the second and third lines intersect corresponds500mL.to the volume of sodium hydroxide consumed by the Human antithrombin III solution: Reconstitute a vial sulfate and the carboxylate groups together (V T).of antithrombin III (see Reagents, Indicators, and Solu-Calculate the molar ratio of sulfate to carboxylate:tions—Reagent Specifications) in water to obtain a solu-tion containing 5 Antithrombin III Units/mL. Dilute this Result = V S/(V T−V S)solution with pH 7.4polyethylene glycol 6000 buffer toobtain a solution having a concentration of 1.0 Anti-Acceptance criteria: The molar ratio of sulfate to car-thrombin III Unit/mL.boxylate is NLT 1.8.。
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eppendorfepTIPS独立包装吸头，50-1000μl，生
物纯级，100个/盒
销售价：293.00元
品牌：艾本德/Eppendorf
订货号：CSS00136
型号规格：德国
产品包装尺寸
长：185mm
宽：145mm
高：90mm
重量：0.5KG
产品参数
类别： 吸头
容量(μl)：
1000
量程(μl)：
50-1000
灭菌： 是
滤芯： 无
描述： 盒装
销售包装数量： 100个/盒
质量、方便、高效的一站式采购平台
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epTIPS 独立包装, 50-1000μl, 生物纯级, 100个吸头