DOTAP DOTAP 哺乳动物细胞转染试剂

Biontex推出的哺乳动物细胞转染试剂DOTAP是一代转染试剂。DOTAP是由单阳离子- -[1-(2,3-二油酰基)]-N,N,N-三甲胺丙烷甲基硫酸盐(C43H83NO8S, M: 774,21)以1mg/ml的浓度在无菌水中制备而成的。大量实验和文献表明DOTAP在体内和体外都适用，虽然DOTAP不能提供现代转染试剂的所有特点，但在大多数情况下都能得出可靠的数据结果。
产品规格
应用                        DNA或者反义核苷酸寡聚体转染哺乳动物细胞   
制定                        单阳离子- -[1-(2,3-二油酰基)]-N,N,N-三甲胺丙烷甲基硫酸盐以1mg/ml的浓度在无菌水中制备而成的   
    检测                        每1ml试剂可使用150次/六孔板、75次/60mm培养皿   
    运输                        常温   
    储存                        40 C   
   保质期                       一年   
目录号
    产品                     货号                           规格                   目录价    
DOTAP                   T010-0.5                        0.5 ml                    1000   
DOTAP                   T010-1.0                        1.0 ml                    2000   
DOTAP                   T010-2.0                       2 ×1.0 ml                 2700   
DOTAP                   T010-5.0                       5× 1.0 ml                 8000 
针对的细胞(参考文献)
Cells successfully transfected with DOTAP (incomplete list)
Cell Line Species & Type References   
A375-C6 human, melanoma Muthukkumar, S. et al, J. Biol. Chem. 271, 1996, 5733
Sells, S.E. et al., Mol. Cell. Biol. 15, 1995, 682   
A549 human, B-cell line Zauner, W. et al., BioTechniques 20, 1996, 905   
B6.NOS mouse, fibroblast, a derivative of B6 fibroblasts Pantopoulos, K. et al.; Proc. Natl. Acad. Sci. USA 92, 1995, 1267   
Balb/3T3 mouse, fibroblast Pickering, J.G. et al., Circulation 89, 1994, 13
Amri, E.-Z. et al., J. Biol. Chem. 270, 1995, 2367
Capaccioli, S. et al., Biochem. Biophys. Res. Comm. 197, 1993, 818   
BalbSV mouse (H-2d) Walker, C. et al., Proc. Natl. Acad. Sci. USA 89, 1992, 7915   
BHK syrian hamster, kidney Bucci, C. et al., Cell 70, 1992, 715   
BNL CL.2   Zauner, W. et al., BioTechniques 20, 1996, 905   
C2 mouse, myoblasts Albagli-Curiel, O. et al., Differentiation 52, 1993, 201
Carnac, C. et al., Oncogene 8, 1993, 3103   
C6   Lowe, W.L. et al.; Biochem. Biophys. Res. Comm. 189, 1992, 972   
Cardiocyte rat, primary neonatal Disch, D.L. et al., Mol. Cell. Biol. 16, 1996, 4043   
Cardiomyoctes mouse, primary Sack, M.N. et al., Proc. Natl. Acad. Sci. 94, 1997, 6438   
CaShi human, cervical cancer Lappalainen, K. et al., Biochimica Biophysica Acta 1196, 1994, 201   
CCRF-CEM/VLB human, leukemic T-lymphoblasts Capaccioli, S. et al., Biochem. Biophys. Res. Comm. 197, 1993, 818   
CHO Chinese hamster ovary Topol, L. Z. et al., Mol. Cell Biol. 17, 8, 1997, 4801
Tsai-Morris, C-H. et al., J. Biol. Chem 270, 1995, 7487   
Clone 9 insect cells, Spodoptera frugiperda Sedlik, C. et al., Proc. Natl. Acad. Sci. 94, 1997, 7503   
COS-1 african green mon
key, kidney, SV 40 transformed Baltensberger, K. et al., Science 260, 1993, 1950
Downes, M. et al., Nucl. Acids Res. 24, 1996, 4379   
COS-7 african green mon
key, kidney, SV 40 transformed Fritz, J.D. et al., Hum. Gene Therapy 7, 1996, 1395
Czech, M.P., et al., J. Cell Biol. 123, 1993, 127
Weber, M. et al., BioTechniques19, 1995, 930
McLachlan, G. et al., Gene Therapy 2, 1995, 614
Zou J.X. et al., Proc. Natl Acad. Sci. USA 93, 1996, 326   
CV-1 african green mon
key, kidney Forman, B.M. et al. Cell 81, 1995, 541 & 687   
D-17 dog, canis familiaris, primary osteogenic sarcoma Willems, L.. et al., Virology 189, 1992, 775   
DTM rat Topol, L. Z. et al., Mol. Cell. Biol. 17, 8, 1997, 4801   
eEnd2 Endothelial cell, derivative from an embryonal hemangioma of a chimeric mouse Besser, D. et al., Oncogene 11, 1995, 2383   
EL-4 mouse, (H-2b), lymphoma Chen, W. et al., J. Immunol. Methods 160, 1993, 49   
ES cells mouse Strauss, W.M. et al., Science 259, 1993, 1904
Lee, J.T. et al., Nucl. Acids Res. 24, 1996, 5054   
ES cells (Dnmt and J1 mutants) mouse Tucker, K.L. et al., Proc. Natl. Acad. Sci. USA 93, 1996, 12920   
Fibroblasts human, primary Zauner, W. et al., BioTechniques 20, 1996, 905   
Fibroblasts Fisher rat embryo fibroblasts Reed, M. et al., Gene Expression 3, 1993, 95   
Fibroblasts, rat dermis, fetal, primary cells Lowe, W.L. et al., Biochem. Biophys. Res. Comm. 189, 1992, 972   
GP+envAm12 amphotropic retrovirus packaging cell line Kume, A. et al., Blood 84, 1994, 3311   
GSM06 mouse, gastric surface mucous cells from transgenic mice Konda, Y. et al., J. Clin. Invest. 99, 8, 1997, 1842   
H225 human, melanoma Zauner, W. et al., BioTechniques 20, 1996, 905   
HEL human, hematopoietic Bastian, L.S. et al., J. Biol. Chem. 271, 1996, 18554
Disch, D.L. et al., Mol. Cell. Biol. 16, 1996, 4043   
HeLa human, cervix, epitheloid carcinoma Fritz, J.D. et al., Hum. Gene Therapy 7, 1996, 1395
Zhang, W.-W. et al., BioTechniques 15, 1993, 868
Weber, M. et al., BioTechniques 19, 1995, 930
Doppler, C. et al., Aids Res. Hum. Retroviruses 8, 1992, 245
Yamanaka, R. et al., Proc. Natl. Acad. Sci 94, 1997, 6462   
HeLa tat-III derivative of HeLa Doppler, C. et al., Aids Res. Hum. Retroviruses 8, 1992, 245   
HeLa TK-  derivative of HeLa Treier, M. et al., Cell 78, 1994, 787   
Hepa 1-6 murine hepatocarcinoma Ghoumari, A.M. et al., Gene Therapy 3, 1996, 483   
Hepatocytes Primary Fisher rat hepatocytes Holmen, S.L. et al., In Vitro 31, 1995, 347   
HepG2 human, hepatocellular carcinoma Ghoumari, A.M. et al., Gene Therapy 3, 1996, 483
Zhang, W.-W. et al., BioTechniques 15, 1993, 868   
Hippocampal neurons mouse, primary cells Marsden, K.M. et al., J. Neurosci. 16, 1996, 3265   
Hippocampal neurons rat, primary cells Kaech, S. et al., Mol. Brain Res. 35, 1996, 344   
HMEC-1 human, microvascular endothelial cells Schmedtje, J.F. et al., Biochem. Biophys. Res. Comm. 220, 1996, 687   
Huh7 human, hepatocarcinoma Weber, M. et al., BioTechniques19, 1995, 930   
I-3 mouse, H-2Kb-transfected L cell Chen, W. et al., J. Immunol. Methods 160, 1993, 49   
I10 mouse, Leydig testicular tumor Tsai-Morris, C-H. et al., J. Biol. Chem 270, 1995, 7487   
JEG-3 human, choriocarcinoma Downes, M. et al., Nucl. Acids Res. 24, 1996, 4379   
Jurkat human, acute T-cell leukemia Doppler, C. et al., Aids Res. Hum. Retroviruses 8, 1992, 245   
LHR expressing Leydig tumor cell line mouse Tsai-Morris, C-H. et al., J. Biol. Chem 270, 1995, 7487   
LH chicken, liver Weber, M. et al., BioTechniques19, 1995, 930   
LHR non-expressing cell line mouse Tsai-Morris, C-H. et al., J. Biol. Chem 270, 1995, 7487   
Ltk mouse, connective tissue Levine, A. et al., Proc. Natl. Acad. Sci. USA 89, 1992, 10119   
LoVo/Dx human, colon adenocarcinoma Capaccioli, S. et al., Biochem. Biophys. Res. Comm. 197, 1993, 818   
Lymphocytes, PBL human, peripheral blood, primary cells Abken, H. et al., Nucleic Acids Res. 20, 1992, 3527
Abken, H. et al., Proc. Natl. Acad. Sci USA 90, 1993, 6518   
M17 human, neuroblastoma Petersen, R.B. et al., J. Biol. Chem. 271, 1996,12661   
Macrophages mouse (C57BL/6 or BALB/c), primary cells Rouse, R.J.D. et al., J. Virol. 68, 1994, 5685   
MC57 mouse (H-2b) Walker, C. et al., Proc. Natl. Acad. Sci. USA 89, 1992, 7915   
MCF-7 human, breast adenocarcinoma, pleural effusion Bertram, J. et al., Biochem Res. Commun. 200,1994, 661
Schräder, M. et al., J. Biol. Chem. 269, 1994, 5501 & 6444
Schräder, M. et al,. J. Mol. Endocrinol. 12, 1994, 327
Schräder, M. et al,. Nature 370, 1994, 382
Schräder, M. et al,. Mol. Cell. Biol. 15, 1995, 1154   
MDA-MB-435 human, ductal carcinoma, breast Lu, D. et al., Cancer Gene Therapy 1, 1994, 245 & 317 Abstract   
MEC-J erythroleukemic, murine Macleod, K.F. et al., Genes, Develop. 9, 1995, 935   
MH1C1 rat, hepatoma cells Perletti, G. et al., Biochem. Biophys. Res. Comm. 221, 1996, 688   
mhAT3F hepatocytic cell line from tumorous liver of transgenic mouse Vallet, V. et al., Mol. Cell. Biol. 15, 1995, 5453   
mhAT3F hepatocyte-like cell line derived from tumorous liver of transgenic mice Rencurel, F. et al., Biochem. J. 314, 1996, 903   
NG 108-15 neuroblastoma(N18TG-2, mouse) x glioma (C6 BV-1, rat) hybrid Pilon, C. et al., Europ. J. Pharmacol. 268, 1994, 129   
NIH/3T3 NIH Swiss mouse, embryo, contact inhibited, Fritz, J.D. et al., Hum. Gene Therapy 7, 1996, 1395
Zauner, W. et al., BioTechniques 20, 1996, 905
Zou, J.X. et al., Proc. Natl. Acad. Sci. USA 93, 1996, 326   
PXF 1118 human, pleural mesothelioma, Kiehntopf, M., et al., EMBO J. 13, 1994, 4645   
RASM rat, primary aortic smooth muscle cells Schmedtje, J.F. et al., Biochem. Biophys. Res. Comm. 220, 1996,687   
REF-1 rat Topol, L. Z. et al., Mol. Cell Biol. 17, 8, 1997, 4801   
Ret-1   Zauner, W. et al., BioTechniques 20, 1996, 905   
ROS 17/2.8 rat, osteosarcoma Kahlen, J.-P. et al., Biochem. Res. Commun. 202, 1994, 13666
Schräder, M. et al,. Mol. Cell. Biol. 15, 1995, 1154   
SCC-13 keratinocytes from epidermoid carcinoma Luchi, S. et al., Proc. Natl. Acad. Sci. 94, 1997, 7948   
SK-BR-3 human, breast adenocarcinoma, malignant, pleural effusion Bertram, J. et al., Biochem. Res. Commun. 200, 1994, 661   
Smooth muscle cells human, internal artery, mammary; primary cells, explant, or organ culture Pickering, J.G. et al., Circulation 89, 1994, 13   
Smooth muscle cells New Zealand White Rabbit, thoracic aorta; primary cells, explant, or organ culture Pickering, J.G. et al., Circulation 89, 1994, 13   
Smooth muscle cells rat, thoracic aorta and vena cava SMC cells, primary Eckhard, A.D. et al., Proc. Natl. Acad. Sci. 94, 1997, 9487   
Spleen cells mouse, C57BL/6 (H-2b); primary cells Chen, W. et al., J. Immunol. Methods 160, 1993, 49   
ST33c rat Topol, L. Z. et al., Mol. Cell Biol. 17, 8, 1997, 4801   
T2-Kb human, lymphoblastoid cell line Chen, W. et al., J. Immunol. Methods 160, 1993, 49   
U87   Garcia, M.A. et al., J. Neurosci. 16, 1996, 7550   
Vero (CCL 81) african green mon
key, kidney Defechereux, P. et al., J. Virol. 67, 1993, 4379
Jackers, P. et al., J. Virol. 66, 1992, 3899   
WI-38 (CCL75) human, embryonic lung fibroblasts Pan, Z.K. et al., J. Clin. Invest. 98, 1996, 2042
Quattrone, A. et al., Cancer Res. 55, 1995, 90   
WI-38 VA-13 human, SV40-transformed derivative of WI-38 cells Quattrone, A. et al., Cancer Res. 55, 1995, 90   
Y1 (Cll 75) mouse, adrenal tumor Tsai-Morris, C-H. et al., J. Biol. Chem 270, 1995, 7487   
293 human, transformed primary embryonal kidney Fritz, J.D. et al., Hum. Gene Therapy 7, 1996, 1395
Kummer-Singh, R. et al. Hum. Mol. Gen. 5, 1996, 913
Roth, J.A et al. Proc. Am. Assoc. Cancer Res. 35, 1994, 692
Zhang, W.-W. et al., BioTechniques 15, 1993, 868 
In vivo applications (incomplete list)
Species / Target Tissue Reference   
Mouse, Balb/c Templeton, N. S. et al., Nature Biotechnology 15, 1997, 647   
Mouse, C57Bl/6(H-2b) Walker, C. et al., Proc. Natl. Acad. Sci. USA 89, 1992, 7915   
Mouse, C57BL/6 male Crespo, J. et al., Biochem. Pharmacol. 51, 1996, 1309   
Mouse, C57BL/6 Marinaro, M. et al., J. Exp. Med. 185, 3, 1997, 415   
Mouse, Swiss-Webster Meiri, N. et al., Proc. Natl. Acad. Sci. 94, 1997, 4430   
Mouse, normal, or CF mutant mouse; Bronchial epithelium, lung parenchyma Alton, E.W.F.W. et al., Nature genetics 5, 1993, 135
McLachlan, G. et al., Gene Therapy 2, 1995, 614   
rat, Sprague-Dawley, male; Kidney Boletta, A. et al., Human Gene Therapy 8, 1997, 1243   
Sheep, Rabbit; Lymphocytes Willems, L. et al., Virology 189, 1992,775   
Chicken, White Leghorn, stage10-15; Primordial germ cells, cells of the blood circulatory system Watanabe, M. et al., Mol. Reprod. Dev. 38, 1994, 268   
NIH mice, male; Skin Alexander, M.Y. et al., Hum. Mol. Genet. 4, 1995, 2279   
Mouse, athymic nude; Tumor xenograft (MDA-MB-435) Lu, D. et al., Cancer Gene Thrapy, 1, 1994, 245 
一代转染试剂
这些试剂一般都是基于单阳离子脂质和季铵离子。没有精确的缓冲性能，也没有内涵体缓冲选择，以此作为比现代转染试剂效率低的理由。另外，大多数转染试剂受血清YZ而DOTAP除外。
这些试剂的另一特点是它们都是进行传统的转染，在转染之前需提供细胞接种和增值的相关信息以及转染之后要进行培养基的更换。为了得到满意的结果必须小心优化DNA和脂质之间的比列已成为一种准则。