碧云天_BCA蛋白浓度测定试剂盒

BCA蛋白浓度测定试剂盒 产品编号 产品名称 包装 P0012 BCA蛋白浓度测定试剂盒 500次 产品简介： ¾ BCA蛋白浓度测定试剂盒(BCA Protein Assay Kit)是根据目前世界上最常用的两种蛋白浓度检测方法之一BCA法研制而 成，实现了蛋白浓度测定的简单，高稳定性，高灵敏度和高兼容性。 ¾ 灵敏度高，检测浓度下限达到25µg/ml，最小检测蛋白量达到0.5µg，待测样品体积为1-20µl。 ¾ 在50-2000µg/ml浓度范围内有较好的线性关系。 ¾ BCA法测定蛋白浓度不受绝大部分样品中的化学物质的影响，可以兼容样品中高达5%的SDS，5%的Triton X-100，5%的 Tween 20, 60, 80。但受螯合剂和略高浓度的还原剂的影响，需确保EDTA低于10mM，无EGTA，二硫苏糖醇低于1mM， β-巯基乙醇低于0.01%。不适用BCA法时建议试用碧云天生产的Bradford蛋白浓度测定试剂盒(P0006)。 ¾ BCA蛋白浓度测定试剂盒对样品中各种物质的详细的兼容性，请参见碧云天如下网页: http://www.beyotime.com/Compatibility Chart For BCA Kit.pdf ¾ 每个试剂盒可以检测500个样品。 包装清单： 产品编号 产品名称 包装 P0012-1 BCA试剂 A 100ml P0012-2 BCA试剂 B 3ml P0012-3 蛋白标准(5mg/ml BSA) 1ml — 说明书 1份 保存条件： BCA试剂A和B室温保存，蛋白标准请-20℃冻存。本试剂盒自订购之日起一年内有效。 ： ¾ 需酶标仪一台，测定波长为540-595nm之间，562nm最佳。需96孔板。如果没有酶标仪，也可以使用普通的分光光度计测 定，但测定时，需根据比色皿的最小检测体积，适当加大BCA工作液的用量使不小于最小检测体积，样品和标准品的用 量可相应按比例放大也可不变。使用分光光度计测定蛋白浓度时，每个试剂盒可以测定的样品数量可能会显著减少。 ¾ 如发现样品稀释液或裂解液本身就有较高背景，请试用碧云天生产的Bradford蛋白浓度测定试剂盒(P0006)。 ¾ 为了加快BCA法测定蛋白浓度的速度可以适当用微波炉加热，但是切勿过热。 ¾ EDTA浓度必须小于10mM，不兼容EGTA。不适用BCA法时，请试用碧云天生产的Bradford蛋白浓度测定试剂盒(P0006)。 ¾ 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. 根据样品数量，按50体积BCA试剂A加1体积BCA试剂B(50:1)配制适量BCA工作液，充分混匀。BCA工作液室温24小时内 稳定。 2. 完全溶解蛋白标准品，取10µl稀释至100µl，使终浓度为0.5mg/ml。蛋白样品在什么溶液中，标准品也宜用什么溶液稀释。 但是为了简便起见，也可以用0.9%NaCl或PBS稀释标准品。 3. 将标准品按0, 1, 2, 4, 8, 12, 16, 20µl加到96孔板的标准品孔中，加用于稀释标准品的溶液补足到20µl。 4. 加适当体积样品到96孔板的样品孔中，加用于稀释标准品的溶液到20µl。 5. 各孔加入200µl BCA工作液，37℃放置30分钟。 注: 也可以室温放置2小时，或60℃放置30分钟。BCA法测定蛋白浓度时，吸光度会随着时间的延长不断加深。并且显色 反应会因温度升高而加快。如果浓度较低，适合在较高温度孵育，或延长孵育时间。 6. 测定A562，540-595nm之间的波长也可接受。根据标准曲线计算出蛋白浓度。 常见问题： 1. 测定标准曲线时发现随着标准品浓度的增加吸光度或颜色没有明显变化。 可能的原因是样品中含有严重干扰BCA法测定蛋白浓度的物质，详细的BCA法的兼容性列表请参考碧云天如下网页: http://www.beyotime.com/Compatibility Chart For BCA Kit.pdf 2. 是否每次测定时都需要做标准曲线？ 建议每次测定时都做标准曲线。因为BCA法测定时颜色会随着时间的延长不断加深，并且显色反应的速度和温度有关， 所以除非精确控制显色反应的时间和温度，否则如需精确测定宜每次都做标准曲线。 使用本产品的文献： 1. Liu MJ, Wang Z, Li HX, Wu RC, Liu YZ, Wu QY. Mitochondrial dysfunction as an early event in the process of apoptosis induced by woodfordin I in human leukemia K562 cells. Toxicol Appl Pharmacol. 2004 Jan 15;194(2):141-55. 2. Wu RC, Chen DF, Liu MJ, Wang Z. Dual effects of cycloheximide on U937 apoptosis induced by its combination with VP-16. Biol Pharm Bull. 2004 Jul;27(7):1075-80. 3. Zheng CH, Gao JQ, Zhang YP, Liang WQ. A protein delivery system: biodegradable alginate-chitosan-poly(lactic-co-glycolic acid) composite microspheres. Biochem Biophys Res Commun. 2004 Oct 29;323(4):1321-7. 4. R.-C. Wu, Z. Wang, M.-J. Liu, D.-F. Chen and X.-S. Yue. β2-integrins mediate a novel form ofchemoresistance in cycloheximide-induced U937 apoptosis. Cell. Mol. Life Sci. 2004,61(16):2071–2082. 5. 刘军 丁劲 薛采芳 李英辉 宫卫东 赵亚 黄豫晓。 带有蛋白转导域的靶向核糖核酸酶对乙型肝炎病毒复制的影响。 中华肝脏病杂志2004年第6期第377页。 6. Zhong-shu Liang , Kan Yang and Ben-mei Chen. Cholesterol in U937 foam cells assayed in liquid chromatographic-mass spectrometry. Chin J Mod Med; 2004, 14(20): 51-56. 7. Liu MJ, Yue PY, Wang Z, Wong RN. Methyl protodioscin induces G2/M arrest and apoptosis in K562 cells with the hyperpolarization of mitochondria. Cancer Lett. 2005, 224:229–241. 8. Liu MJ, Wang Z, Ju Y, Wong RN, Wu QY. Diosgenin induces cell cycle arrest and apoptosis in human leukemia K562 cells with the disruption of Ca2+ homeostasis. Cancer Chemother Pharmacol. 2005 Jan;55(1):79-90. 9. Jin Ding, Jun Liu, Cai-Fang Xue, Ying-Hui Li, Ya Zhao, Jun Chen, Yu-Xiao Huang, Zhong-Xiang Liu. TatPTD can introduce HBV targeted ribonuclease into hepatocytes. World Chin J Digestol. 2005 april 15;13(8):958-962. 10.Zhao Y, Guan H, Liu SF, Wu RC, Wang Z. Overexpression of QM induces cell differentiation and mineralization in MC3T3-E1. Biol Pharm Bull. 2005 Aug;28(8):1371-6. 11.Le-Feng Zhang, Shuang-Qing Peng, Sheng Wang. Influence of lead (Pb2+) on the reactions of in vitro cultured rat aorta to 5-hydroxytryptamine. Toxicology Letters 159 (2005) 71–82. 12.Jin Ding, Jun Liu, Cai-Fang Xue, Ying-Hui Li, Ya Zhao, Jun Chen, Yu-Xiao Huang, Zhong-Xiang Liu TatPTD can introduce HBV targeted ribonuclease into hepatocytes. Shijie Huaren Xiaohua Zazhi 2005;13(8):958-962 13.Gao Jianmei, Wan Yanzhen, Han Jun, Wang Xiaofan, Chen Lan, Nie Kai, Zhou Wei, Dong Xiaoping. Influence of the Numbers of Octapeptide Repeats within N2terminus of Recombinant Human PrP Proteins on the Protease Resistance af ter Interacting with Metal Ions and the Binding Abil ity with Tau Protein. Chinese Journal of Virology. 21(5):376-83. 14.Wang Yichao, Zou Quanming, Ren Jianmin, Liu Jian,Wang Xiaoqin. Preparation and character of microspheres of Helicobacter pylori whole cell protein encapsulated by chitosan - alginate. West China Journal of Pharmaceutical Sciences. 2005,20(5):375-7. 15.Wu ZQ, Li M, Chen J, Chi ZQ, Liu JG. Involvement of cAMP/cAMP-dependent protein kinase signaling pathway in regulation of Na+,K+-ATPase upon activation of opioid receptors by morphine. Mol Pharmacol. 2006 Mar;69(3):866-76. 16.Yi ZC, Liu YZ, Li HX, Yin Y, Zhuang FY, Fan YB, Wang Z. Tellimagrandin I enhances gap junctional communication and attenuates the tumor phenotype of human cervical carcinoma HeLa cells in vitro. Cancer Lett. 2006 Oct 8;242(1):77-87. 17.Ma JF, Guo JK, Peng LW, Chen CY, Zhang LX. Decrease of photosystem II photochemistry in Arabidopsis 1 mutant is dependent on leaf age. J Integr Plant Biol. 2006; 48(12), 1409-1414. 18.Huang YY, Deng JY, Gu J, Zhang ZP, Maxwell A, Bi LJ, Chen YY, Zhou YF, Yu ZN, Zhang XE. The key DNA-binding residues in the C-terminal domain of Mycobacterium tuberculosis DNA gyrase A subunit (GyrA). Nucleic Acids Res. 2006;34(19):5650-9. 19.Ma JF, Guo JK, Peng LW, Chen CY, Zhang LX. Decrease of Photosystem II Photochemistry in Arabidopsis ppt1 Mutant Is Dependent on Leaf Age. Journal of Integrative Plant Biology. 2006,48(12):1409-1414. 20.Ji G, Wang M, Long AH, Liu CH. Regulative Effects of"Qinggan Huoxue Recipe" and Its Separated Recipes on Kupffer Cells Mediated by LPS. Shanghai Chinese Medicine magazine.2007 Jan;Vol.41 No.1. 21.Li JL, Yan ZP, Chen XH, Wang YJ, Sun M, Shi YX, Wang CB. Inh ib ition of Polypeptide from Chlamys fa rreri on UVA2Induced Apoptosis of HaCaT Cells Depending on p38MAPK Pa thway and Ca spase23. Chin Pharm J.2007 January, Vol.42 No.2. 22.Wu ZQ, Chen J, Chi ZQ, Liu JG. Involvement of dopamine system in regulation of Na+,K+-ATPase in the striatum upon activation of opioid receptors by morphine. Mol Pharmacol. 2007 Feb;71(2):519-30. 23.Zong-Chun Yi, Hong Wang, Guang-Yao Zhang, Bing Xia. Downregulation of connexin 43 in nasopharyngeal carcinoma cells is related to promoter methylation. Oral Oncol. 2007 Feb 14; 24.Gao J, Zhao YN, Jiang QW. Investigation of Asymptomatic Infection of Echovirus 30 in Population of Shanghai. Journal of the virus.2007 March;Vol.23 No.2. 25.Deng XQ, Chen LL, Li NX. The expression of SIRT1 in nonalcoholic fatty liver disease induced by high-fat diet in rats. Liver Int. 2007 Jun;27(5):708-15. 26.Gao MQ, Guo SB, Chen XH, Du W, Wang CB. Molecular mechanisms of polypeptide from Chlamys farreri protecting HaCaT cells from apoptosis induced by UVA plus UVB. Acta Pharmacol Sin. 2007 Jul;28(7):1007-14. 27.Dong S, Li C, Wu P, Tsien JZ, Hu Y. Environment enrichment rescues the neurodegenerative phenotypes in presenilins-deficient mice. Eur J Neurosci. 2007 Jul;26(1):101-12. 28.Wang Q, Liang C, Song J, Chen X. HA2 from the Helicoverpa armigera nucleopolyhedrovirus: a WASP-related protein that activates Arp2/3-induced actin filament formation. Virus Res. 2007 Jul;127(1):81-7. 29.Wang YC, Zou QM, Ren JM, Guo G, Xie QH, Liu j. Study of the microspheres technology of Helicobacter pylori whole cell protein encapsulated by chitosan2alginate. Chongqing Medical.2007 July;Vol.36 No.13. 30.Li JL, Liu N, Chen XH, Sun M, Wang CB. Inhibition of UVA-induced apoptotic signaling pathway by polypeptide from Chlamys farreri in human HaCaT keratinocytes. Radiat Environ Biophys. 2007 Aug;46(3):263-8. 31.Yuan QL, Yang CX, Xu P, Gao XQ, Deng L, Chen P, Sun ZL, Chen QY. Neuroprotective effects of ginsenoside Rb1 on transient cerebral ischemia in rats. Brain Res. 2007 Sep 5;1167:1-12. 32.Chen YR, Nie SD, Shan W, Jiang DJ, Shi RZ, Zhou Z, Guo R, Zhang Z, Li YJ. Decrease in endogenous CGRP release in nitroglycerin tolerance: role of ALDH-2. Eur J Pharmacol. 2007 Sep 24;571(1):44-50. 33.Yi ZC, Wang H, Zhang GY, Xia B. Downregulation of connexin 43 in nasopharyngeal carcinoma cells is related to promoter methylation. Oral Oncol. 2007 Oct;43(9):898-904. 34.Wang ZL, Li JL, Wang CB. The role of tumor necrosis factor rela ted apoptosis induc ing ligand and Polypeptide from Chlam ys fa rreri in UVA2induced apoptosis in HaCaT cells. Chinese Pharm acological B ulletin.2007 Oct;23(10):1375～9. 35.Zhang Y, Zhen DX, Ren TL, Wang DP. The Character istics Effects of Ga tifloxac in on L iverM icrosome Cytochrome P450 in Rats. Pharm J Chin PLA.2007 Dec;Vol.23 NO.6. 36.Pan YQ, Ding D, Li DX, Chen SQ. Expression and bioactivity analysis of Staphylococcal enterotoxin M and N. Protein Expr Purif. 2007 Dec;56(2):286-92. 37.Jun Hong, Dongmei Xua, Peijun Gong, Hanwen Suna, Li Dong, Side Yao. � Covalent binding of -chymotrypsin on the magnetic nanogels covered by amino groups. Journal of Molecular Catalysis B: Enzymatic 45 (2007) 84–90. 38.Wang YL, Wang CY, Zhang BJ, Zhang ZZ. Shenfu injection suppresses apoptosis by regulation of Bcl-2 and caspase-3 during hypoxia/reoxygenation in neonatal rat cardiomyocytes in vitro. Mol Biol Rep. 2007 Nov 30. 39.Huang JW, Tian LX, Du ZY, Yang HJ, Liu YJ. Effects of dietary thiamin on the physiological status of the grouper Epinephelus coioides. Fish Physiol Biochem. (2007) 33:167–172. 40.Cheng BQ, Jiang B, Bao J. Construction and expression of ga lectin2 3 shRNA recombinant vector and its effect on prolifera tion ra te of colorecta l cancer lovo cells. J Fourth MilMedUniv.2007，28(11)。 41.Wang YC, Zou QM, Mao XH, Guo G, Xie QH. A prelim inary non2clin ica l study of Helicobacter pylori m icrospheres vacc ine. Chinese Journal of New Drugs.2007,Vol.16 No.7. 42.Wang M, Long AH, Liu CH, Ji G. Effect of Qinggan Huoxue recipe and its separated recipe on ERK signal ing pathway in Kupffer cell mediated by LPS. Integrative Medicine in the Journal of liver disease.2007;Vol.17 No.5. 43.Nie H, Li Z, Lukas RJ, Shen Y, Song L, Wang X, Yin M. Construction of SH-EP1-alpha4beta2-hAPP695 Cell Line and Effects of Nicotinic Agonists on beta-amyloid in the Cells. Cell Mol Neurobiol. 2008 Jan;28(1):103-12. 44.Zhang DW, Cheng Y, Wang NL, Zhang JC, Yang MS, Yao XS. Effects of total flavonoids and flavonol glycosides from Epimedium koreanum Nakai on the proliferation and differentiation of primary osteoblasts. Phytomedicine. 2008 Jan;15(1-2):55-61. 45.Zhang H, Deng JY, Bi LJ, Zhou YF, Zhang ZP, Zhang CG, Zhang Y, Zhang XE. Characterization of Mycobacterium tuberculosis nicotinamidase/pyrazinamidase. FEBS J. 2008 Feb;275(4):753-62. 46.Zhang GG, Bai YP, Chen MF, Shi RZ, Jiang DJ, Fu QM, Tan GS, Li YJ. Asymmetric dimethylarginine induces TNF-alpha production via ROS/NF-kappaB dependent pathway in human monocytic cells and the inhibitory effect of reinioside C. Vascul Pharmacol. 2008 Feb-Mar;48(2-3):115-21. 47.Yu S, Geng J, Zhou P, Wang J, Feng A, Chen X, Tong H, Hu J. Application of a new hybrid organic-inorganic monolithic column for efficient deoxyribonucleic acid purification. Anal Chim Acta. 2008 Mar 24;611(2):173-81. 48.Zhang GG, Shi RZ, Jiang DJ, Chen YR, Jia-Chen , Tang ZY, Bai YP, Xiao HB, Li YJ. Involvement of the endothelial DDAH/ADMA pathway in nitroglycerin tolerance: the role of ALDH-2. Life Sci. 2008 Mar 26;82(13-14):699-707. 49.Li J, Liu W, Ding S, Xu W, Guan Y, Zhang JH, Sun X. Hyperbaric oxygen preconditioning induces tolerance against brain ischemia-reperfusion injury by upregulation of antioxidant enzymes in rats. Brain Res. 2008 May 19;1210:223-9. 50.Fu Y, Zhang Z, Zhang G, Liu Y, Cao Y, Yu J, Hu J, Yin X. Adenovirus-mediated gene transfer of tissue factor pathway inhibitor induces apoptosis in vascular smooth muscle cells. Apoptosis. 2008 May;13(5):634-40. 51.Tian B, Wang Z, Zhao Y, Wang D, Li Y, Ma L, Li X, Li J, Xiao N, Tian J, Rodriguez R. Effects of curcumin on bladder cancer cells and development of urothelial tumors in a rat bladder carcinogenesis model. Cancer Lett. 2008 Jun 18;264(2):299-308. 52.Yuan Q, Li PD, Li BH, Yang XZ, Xu SB, Liu XH, Zhou FX, Zhang WJ. Differential IL-4/Stat6 activities correlate with differential expression of regulatory genes SOCS-1, SHP-1, and PP2A in colon cancer cells. J Cancer Res Clin Oncol. 2008 Jun 7. 53.Wan F, You J, Sun Y, Zhang XG, Cui FD, Du YZ, Yuan H, Hu FQ. Studies on PEG-modified SLNs loading vinorelbine bitartrate (I): preparation and evaluation in vitro. Int J Pharm. 2008 Jul 9;359(1-2):104-10. 54.Tian SL, Wang XY, Ding GH. Repeated electro-acupuncture attenuates chronic visceral hypersensitivity and spinal cord NMDA receptor phosphorylation in a rat irritable bowel syndrome model. Life Sci. 2008 Jul 22. 55.Chen M, Li Y, Yang T, Wang Y, Bai Y, Xie X. ADMA induces monocyte adhesion via activation of chemokine receptors in cultured THP-1 cells. Cytokine. 2008 Aug;43(2):149-59. 56.Wei Y, Weng D, Li F, Zou X, Young DO, Ji J, Shen P. Involvement of JNK regulation in oxidative stress-mediated murine liver injury by microcystin-LR. Apoptosis. 2008 Aug;13(8):1031-42.