磷酸钙法细胞转染试剂盒
收藏
产品编号 C0508
数量
¥ 292.00
产品包装:
>200次
产品简介
使用说明
相关文件下载
相关产品
相关论文
产品问答

碧云天生产的磷酸钙法细胞转染试剂盒(Calcium Phosphate Cell Transfection Kit),在传统的磷酸钙细胞转染方法的基础上进行了改良,提高了转染效率,并降低了毒性。用磷酸钙法转染细胞,不仅可以瞬时表达,也可以筛选稳定株。
HEK293或HEK293T细胞,即通常所谓的293细胞,是最适合磷酸钙法转染的细胞之一,优化条件后转染效率可以高达90%以上;通常很容易达到40-50%左右的转染效率。大多数常见的细胞例如Hela细胞、CHO细胞等也都适合磷酸钙法转染,但效率比293细胞要略低一些。
本试剂盒不仅适合于大多数贴壁细胞的转染,也适用于一些悬浮细胞的转染。
本试剂盒提供的试剂都经无菌处理,可以直接使用。
关于不同的细胞转染试剂的主要特点和差异方面的比较,以及如何选择适当的细胞转染试剂,可参考碧云天的相关网
页:http://www.beyotime.com/support/transfection.htm
本试剂盒足够转染不少于200个细胞样品。
包装清单:

产品编号 产品名称 包装
C0508-1 BBS溶液 20ml
C0508-2 氯化钙溶液 20ml
说明书 1份

保存条件:
-20℃保存,一年有效。
注意事项:
转染时,把DNA-氯化钙溶液加入到BBS溶液中,不要把BBS溶液加入到DNA-氯化钙溶液中。
高纯度的质粒是获得高转染效率的必要条件,要确保A260/A280大于1.8,并且电泳检测抽提到的质粒90%以上都是超螺旋。
在用磷酸钙法转染细胞时应使用浓度不高于5%的二氧化碳,二氧化碳的最佳浓度为3%左右。
BBS溶液的pH值直接关系到转染效率,尽量避免把BBS溶液长时间暴露在空气中,以免被空气中的二氧化碳酸化。
转染时由于质粒不同、细胞不同,最佳的质粒用量需自行摸索。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
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. 对于贴壁细胞:
a) 将细胞培养于培养皿或培养板内,通常在铺板后1-2天内长到70-80%满为宜。后续说明都按6孔板内一个孔进行说明,如果有更多个孔或大小不相同的培养器皿,请自行换算。
b) 在转染前30-60分钟,吸去细胞培养液,加入新鲜的不含抗生素的完全培养液2毫升。注意:转染时最好使用新鲜配制且pH值经过精心调校的培养液;转染用的培养液可以在配制后分装冻存,使用时再解冻。
c) 取2-6微克待转染的质粒DNA (质粒总体积不宜超过20微升),加入到100微升氯化钙溶液中,混匀。
d) 把DNA-氯化钙溶液加入到100微升BBS溶液中,混匀,室温孵育10-20分钟。此时不会产生可见的沉淀。
e) 把DNA-氯化钙-BBS混合物均匀滴加到整个6孔板内。在含5%二氧化碳的37℃细胞培养箱内培养。
f) 根据实验要求和磷酸钙对于不同细胞的毒性不同,在4-16小时后轻轻晃动培养板数次以充分悬浮一些磷酸钙沉淀,吸去含磷酸钙沉淀的培养液,加入2毫升新鲜的完全培养液,继续培养。
g) 通常在转染约24小时后就可以检测到转染基因的表达。
2. 对于悬浮细胞:
a) 离心收集悬浮细胞,用PBS洗涤一次。
b) 按照上面的步骤c)和d)制备DNA-氯化钙-BBS混合物。
c) 每106个细胞的沉淀,用100微升DNA-氯化钙-BBS混合物重新悬浮,室温放置20分钟。
d) 在一个6孔板孔内加入2毫升完全培养液,然后加入来自上一步的细胞-DNA-氯化钙-BBS混合物,混匀。
e) 在含5%二氧化碳的37℃细胞培养箱内培养。
f) 根据实验要求和磷酸钙对于不同细胞的毒性不同,在4-16小时后离心收集细胞,用PBS洗一次,然后用2毫升完全培养液重新悬浮细胞,继续培养。
g) 通常在转染约24小时后就可以检测到转染基因的表达。
相关产品请点击如下按钮:
使用本产品的文献:
1. Hu K, Gu Y, Lou L, Liu L, Hu Y, Wang B, Luo Y, Shi J, Yu X, Huang H.
J Hematol Oncol 2015 Jan 27;8:1. (IF 11.059)
2. Xu W, Hua H, Chiu YH, Li G, Zhi H, Yu Z, Ren F, Luo Y, Cui W
Cell Rep. 29(5):1311-1322.e5. (IF 8.109)
3. Zheng LH, Bao YL, Wu Y, Yu CL, Meng X, Li YX.
Cancer Lett 2008 Dec 8;272(1):102-9. (IF 7.36)
4. Han L, Wang W, Lu J, Kong F, Ma G, Zhu Y, Zhao D, Zhu J, Shuai W, Zhou Q, Chen P, Ye L, Tao J, Ahmad S, Li F, Sun J.
Cancer Lett 2014 Nov 28;354(2):398-406. (IF 7.36)
5. Shuhua Shan, Jinping Niu, Ruopeng Yin, Jiangying Shi, Lizhen Zhang, Caihong Wu, Hanqing Li, Zhuoyu Li
Acta Pharm Sin B. doi: 10.1016/j.apsb.2021.10.004. (IF 7.097)
6. Zhang J, Cao H, Xie J, Fan C, Xie Y, He X, Liao M, Zhang S, Wang H
Cell Death Dis 2018 Feb 14;9(2):224. (IF 6.304)
7. Qiong Mo, Zhao Xu, Fei Deng, Hualin Wang, Yun-Jia Ning
PLoS Pathog. doi: 10.1371/journal.ppat.1009129. (IF 6.218)
8. Xu X, Shi X, Chen Y, Zhou T, Wang J, Xu X, Chen L, Hu L, Shen X
Metabolism 2018 Aug;85:126-138. (IF 6.159)
9. Zhiyuan Niu, Guanxu Chen, Wei Chang, Pengyang Sun, Zhixia Luo, Huiyong Zhang, Lingtong Zhi, Changjiang Guo, Han Chen, Meichen Yin, Wuling Zhu
J Pathol. doi: 10.1002/path.5585. (IF 5.979)
10. Shangping Xing, Feifei Nong, Yaqin Wang, Da Huang, Jialiang Qin, Yu-Fei Chen, Dan-Hua He, Pei-En Wu, Huicai Huang, Ruoting Zhan, Hui Xu, Yong-Qiang Liu
Pharmacol Res. doi: 10.1016/j.phrs.2022.106059. (IF 5.893)
11. Ma K, Li F, Tang Q, Liang P, Liu Y, Zhang B, Gao X
Chemosphere. 219:961-970. (IF 5.778)
12. Ashfaque Ahmed, Meng Wang, Gaber Bergant, Reza Maroofian, Rongjuan Zhao, Majid Alfadhel, Marwan Nashabat, Muhammad Talal AlRifai, Wafaa Eyaid, Abdulrahman Alswaid, Christian Beetz, Yan Qin, Tengfei Zhu, Qi Tian, Lu Xia, Huidan Wu, Lu Shen, Shanshan Dong, Xinyi Yang, Cenying Liu, Linya Ma, Qiumeng Zhang, Rizwan Khan, Abid Ali Shah, Jifeng Guo, Beisha Tang, Lea Leonardis, Karin Writzl, Borut Peterlin, Hui Guo, Sajid Malik, Kun Xia, Zhengmao Hu
Hum Genet. doi: 10.1007/s00439-020-02226-3. (IF 5.743)
13. Bin Zhu, Linhong Li, Rui Wei, Pei Liang, Xiwu Gao
PLoS Genet. doi: 10.1371/journal.pgen.1009888. (IF 5.174)
14. Zhang S, An S, Hoover K, Li Z, Li X, Liu X, Shen Z, Fang H, Ros VID, Zhang Q, Liu X.
Mol Ecol 2018 Jan;27(2):459-475. (IF 5.163)
15. Zhao Li, Kaikai Mao, Ruoheng Jin, Tingwei Cai, Yao Qin, Yunhua Zhang, Shun He, Kangsheng Ma, Hu Wan, Xuexiang Ren, Jianhong Li
Int J Biol Macromol. doi: 10.1016/j.ijbiomac.2022.07.096. (IF 5.162)
16. Cao X, Deng W, Wei Y, Su W, Yang Y, Wei Y, Yu J, Xu X.
INT J NANOMED 2011;6:3335-49. (IF 5.115)
17. Zhao W, Tan J, Zhu T, Ou J, Li Y, Shen L, Wu H, Han L, Liu Y, Jia X, Bai T, Li H, Ke X, Zhao J, Zou X, Hu Z, Guo H, Xia K
J Genet Genomics. 46(5):247-257. (IF 5.065)
18. Zhou R, Lin ZH, Jiang CS, Gong JX, Chen LL, Guo YW, Shen X.
Acta Pharmacol Sin 2013 Oct;34(10):1325-36. (IF 5.064)
19. Qiu-Ying Yan, Jian-Lu Lv, Xing-Yi Shen, Xing-Nan Ou-Yang, Juan-Zhen Yang, Rui-Fang Nie, Jian Lu, Yu-Jie Huang, Jia-Ying Wang, Xu Shen
Acta Pharmacol Sin. doi: 10.1038/s41401-021-00857-4. (IF 5.064)
20. Dan-Yang Zhu, Jian Lu, Rui Xu, Juan-Zhen Yang, Xiang-Rui Meng, Xing-Nan Ou-Yang, Qiu-Ying Yan, Rui-Fang Nie, Tong Zhao, Yi-di Chen, Yin Lu, Yi-Nan Zhang, Wen-Jun Li, Xu Shen
Acta Pharmacol Sin. doi: 10.1038/s41401-022-00884-9. (IF 5.064)
21. Deng HP, Zhuang R, Jia W, Zhang Y, Zhang Y, Hu Y, Zhang XH, Jin BQ.
J IMMUNOL METHODS 2007 Sep;23(5):499-505. (IF 1.901)
22. Shi JF, Li A, Rao GZ, Li Q.
Shaanxi Journal of Medicine 2008 Jan;37(1):73-9.
24. Yang N, Gong F, Sun L, Yang D, Han X, Ma C, Sun Y.
J Cell Biochem 2010 Aug 1;110(5):1208-18. (IF 4.237)
25. Sun BT, Zheng LH, Bao YL, Yu CL, Wu Y, Meng XY, Li YX.
Eur J Pharmacol 2011 Mar 5;654(2):129-34. (IF 3.263)
27. Song ZB, Bao YL, Zhang Y, Mi XG, Wu P, Wu Y, Yu CL, Sun Y, Zheng LH, Huang YX, Liu B, Li YX.
Biochem J 2011 Jun 1;436(2):457-67. (IF 4.097)
28. Shi Y, Bao YL, Wu Y, Yu CL, Huang YX, Sun Y, Zheng LH, Li YX.
J Biomol Screen 2011 Jun;16(5):525-35. (IF 2.297)
29. Zhong XY, Yu JH, Zhang WG, Wang ZD, Dong Q, Tai S, Cui YF, Li H.
Gene 2012 Feb 1;493(1):44-51. (IF 2.984)
30. Qin L, Sun Y, Liu P, Wang Q, Han B, Duan Y.
J BIOMAT SCI-POLYM E 2013;24(15):1757-66. (IF 2.69)
31. Wang L, Hou Y, Quan H, Xu W, Bao Y, Li Y, Fu Y, Zou S.
Protein Sci 2013 Aug;22(8):1060-70. (IF 3.876)
32. Zhou R, Yao X, Xu X, Wang G, Zhu Z, Chen J, Chen L, Shen X.
Steroids 2013 Oct;78(10):987-95. (IF 1.948)
33. Zhuang J, Zhang H, Zhou R, Chen L, Chen J, Shen X.
PROSTAG OTH LIPID M 2013 Oct;106:45-52. (IF 2.283)
34. Zhang WG, Chen L, Dong Q, He J, Zhao HD, Li FL, Li H.
Gene 2013 Dec 1;531(2):235-42. (IF 2.984)
35. Mi XG, Song ZB, Wu P, Zhang YW, Sun LG, Bao YL, Zhang Y, Zheng LH, Sun Y, Yu CL, Wu Y, Wang GN, Li YX.
Toxicol Lett 2014 Jan 30;224(3):349-55. (IF 3.569)
36. Yang P, Li Z, Fu R, Wu H, Li Z.
Cell Signal 2014 Sep;26(9):1853-62. (IF 3.968)
37. Song ZB, Liu B, Li YY, Wu P, Bao YL, Huang YX, Wu Y, Sun LG, Yu CL, Sun Y, Zheng LH, Li YX.
Cell Signal 2014 Oct;26(10):2266-75. (IF 3.968)
40. Sun Y, Fan J, Zhu Z, Guo X, Zhou T, Duan W, Shen X.
Eur J Pharmacol 2015 Sep 5;762:202-13. (IF 3.263)
41. Li X, Guo L, Zhou X, Gao X, Liang P.
SCI REP-UK 2015 Sep 15;5:14095. (IF 3.998)
42. Cheng Y, Zhou M, Wang Y.
J RECEPT SIG TRANSD 2015 Oct 8:1-8. (IF 1.466)
43. Meng SF, Mao WP, Wang F, Liu XQ, Shao LL.
J Appl Toxicol 2015 Nov;35(11):1398-405. (IF 2.997)
45. Qin HS, Yu PP, Sun Y, Wang DF, Deng XF, Bao YL, Song J, Sun LG, Song ZB, Li YX .
Mol Med Rep 2016 Jun;13(6):5118-24. (IF 2.1)
46. Peng T, Pan Y, Gao X, Xi J, Zhang L, Ma K, Wu Y, Zhang J, Shang Q.
INSECT BIOCHEM MOLEC 2016 Aug;75:89-97. (IF 3.827)
47. Wei X, Zheng C, Peng T, Pan Y, Xi J, Chen X, Zhang J, Yang S, Gao X, Shang Q.
INSECT BIOCHEM MOLEC 2016 Oct 27. pii: S0965-1748(16)30158-8. (IF 3.827)
48. Shi X, Yang J, Cui X, Huang J, Long Z, Zhou Y, Liu P, Tao L, Ruan Z, Xiao B, Zhang W, Li D, Dai K, Mao J, Xi X.
PLoS One 2016 Nov 16;11(11):e0166136. (IF 2.74)
49. Wu Z, Wang L, Xu X, Lin G, Mao H, Ran X, Zhang T, Huang K, Wang H, Huang Q, Xu Q, Hu C.
Mol Immunol 2017 May;85:273-282. (IF 3.641)
50. Zhang T, Li D, Wan L, Chen X, Wang X, Zhong B, Wu Z, Mao H, Hu C.
FISH SHELLFISH IMMUN 2017 Nov;70:568-574. (IF 3.298)
51. Ai HH, Zhou ZL, Sun LG, Yang MT, Li W, Yu CL, Song ZB, Huang YX, Wu Y, Liu L, Yang XG, Zhao YQ, Bao YL, Li YX.
Apoptosis 2017 Nov;22(11):1404-1418. (IF 4.543)
52. Wang L, Wu Z, Huang Q, Huang K, Qi G, Wu C, Mao H, Xu X, Wang H, Hu C.
Dev Comp Immunol 2018 Jan;78:26-34. (IF 3.192)
53. Wang L, Wu Z, Huang Q, Huang K, Qi G, Wu C, Mao H, Xu X, Wang H, Hu C
Dev Comp Immunol 2018 Jan;78:26-34. (IF 3.192)
54. Yang M, Zhao H, Ai H, Zhu H, Wang S, Bao Y, Li Y
Eur J Pharmacol 2018 Jun 5;828:60-66. (IF 3.263)
55. Li G, Zhou R, Zhao X, Liu R, Ye C.
Int J Mol Med. 42(2):2972. (IF 3.098)
56. Qin H, Li W, Sun Y, Bao Y, Sun L, Song Z, Zheng L, Zhao Y, Li Y
Eur J Pharmacol 2018 Oct 5;836:75-82. (IF 3.263)
57. Song D, Yang Q, Lang Y, Wen Z, Xie Z, Zheng D, Yan T, Deng Y, Nakanishi H, Quan Z, Qing H
Neurobiol Learn Mem 2018 Nov;155:435-445. (IF 2.768)
58. Du T, Yang L, Xu X, Shi X, Xu X, Lu J, Lv J, Huang X, Chen J, Wang H, Ye JM, Hu L, Shen X
J Endocrinol 2018 Nov 1. pii: JOE-18-0432.R1. (IF 4.041)
59. Wu H, Liu HY, Liu WJ, Shi YL, Bao D
J Cell Biochem 2018 Nov 28. (IF 4.237)
61. Chen X, Han K, Lin G, Liu C, Wang S, Shi X, Hu Z, Wu C, Xu X, Hu C
FISH SHELLFISH IMMUN. 91:194-201. (IF 3.298)
62. Guo C, Wang X, Zhang H, Zhi L, Lv T, Li M, Lu C, Zhu W
Mol Immunol. 114:108-113. (IF 3.641)
63. Xu K, Xie X, Qi G, Weng P, Hu Z, Han K, Yuan H, Li Y, Ran X, Lin G, Hu C
Dev Comp Immunol. 99:103410. (IF 3.192)
64. Sun X, Hu F, Hou Z, Chen Q, Lan J, Luo X, Wang G, Hu J, Cao Z
Exp Cell Res. 383(1):111495. (IF 3.383)
65. Yinjun Fan, Xiuxia Li, Abd Allah A H Mohammed, Ying Liu, Xiwu Gao
BMC Genomics. doi: 10.1186/s12864-020-6466-7. (IF 3.594)
66. Li X, Ren X, Liu Y, Smagghe G, Liang P, Gao X
PESTIC BIOCHEM PHYS. 163:235-240. (IF 2.751)
67. Bin Zhu, Xi Sun, Ximan Nie, Pei Liang, Xiwu Gao
INSECT BIOCHEM MOLEC. doi: 10.1016/j.ibmb.2019.103283. (IF 3.827)
68. Xin Chen, Kun Han, Tao Zhang, Guoqin Qi, Zeyin Jiang, Chengyu Hu
Fish Physiol Biochem. doi: 10.1007/s10695-019-00729-z. (IF 2.242)
69. Xiaomei Li, Yingshou Lei, Yang Yu, Yaqian Zhang, Wenfeng Zhang, Han Shen, Changli Tao, Fenglin Wu, Shulin Huang, Hongwei Shao
INT J CLIN EXP PATHO. (IF 0.252)
70. Xiaofen Xie, Kang Xu, Huiling Mao, Yangfeng Lv, Panwei Weng, Kaile Chang, Gang Lin, Chengyu Hu
FISH SHELLFISH IMMUN. doi: 10.1016/j.fsi.2020.04.014. (IF 3.298)
71. Zhong-Jian Shen, Yan-Jun Liu, Fang Zhu, Li-Mei Cai, Xiao-Ming Liu, Zhi-Qiang Tian, Jie Cheng, Zhen Li, Xiao-Xia Liu
INSECT BIOCHEM MOLEC. doi: 10.1016/j.ibmb.2020.103391. (IF 3.827)
72. Xinlei Yao, Kun Li, Chen Liang, Zilong Zhou, Jiao Wang, Shuyue Wang, Lei Liu, Chun-Lei Yu, Zhen-Bo Song, Yong-Li Bao, Li-Hua Zheng, Ying Sun, Guannan Wang, Yanxin Huang, Jingwen Yi, Luguo Sun, Yuxin Li
J Biol Chem. doi: 10.1074/jbc.RA120.012849. (IF 4.238)
73. Xin Xu, Yidi Chen, Danyang Zhu, Tong Zhao, Rui Xu, Jiaying Wang, Lihong Hu, Xu Shen
AGING-US. doi: 10.18632/aging.202275. (IF 4.831)
74. Yangfeng Lv, Hang Deng, Yapeng Liu, Kaile Chang, Hailing Du, Pengcheng Zhou, Huiling Mao, Chengyu Hu
Dev Comp Immunol. doi: 10.1016/j.dci.2020.103834. (IF 3.192)
75. Hang Deng, Liugen Zeng, Kaile Chang, Yangfeng Lv, Hailing Du, Shina Lu, Yapeng Liu, Pengcheng Zhou, Huiling Mao, Chengyu Hu
Dev Comp Immunol. doi: 10.1016/j.dci.2021.104014. (IF 3.192)
76. Qiong-Ying Hu, Yun Yuan, Yu-Chen Li, Lu-Yao Yang, Xiang-Yu Zhou, Da-Qian Xiong, Zi-Yi Zhao
Biomed Res Int. doi: 10.1155/2021/5574282. (IF 2.276)
77. Dan-Hua He, Yu-Fei Chen, Yi-Le Zhou, Shi-Bing Zhang, Ming Hong, Xianjun Yu, Su-Fen Wei, Xiang-Zhen Fan, Si-Yi Li, Qi Wang, Yongzhi Lu, Yong-Qiang Liu
Cancer Sci. doi: 10.1111/cas.15005. (IF 4.966)
78. Shina Lu, Xiaojue Peng, Gang Lin, Kang Xu, Shanghong Wang, Weihua Qiu, Hailing Du, Kaile Chang, Yangfeng Lv, Yapeng Liu, Hang Deng, Chengyu Hu, Xiaowen Xu
Fish Shellfish Immunol. doi: 10.1016/j.fsi.2021.07.005.
79. Kaikai Mao, Ruoheng Jin, Zhijie Ren, Junjie Zhang, Zhao Li, Shun He, Kangsheng Ma, Hu Wan, Jianhong Li
Insect Sci. doi: 10.1111/1744-7917.12910. (IF 2.791)
80. Yang Cheng, Tengfei Lu, Junliang Guo, Zhe Lin, Qiao Jin, Xiaoming Zhang, Zhen Zou
Open Biol. doi: 10.1098/rsob.210307. (IF 4.931)
81. Xiaodong Li, Yichang Wang, Wei Wu, Jianyang Xiang, Lei Qi, Ning Wang, Maode Wang, Hai Yu
J Oncol. doi: 10.1155/2022/4966820. (IF 2.206)
82. Meng-Yuan Zhang, Pei Zhang, Xu Su, Tian-Xin Guo, Jun-Lei Zhou, Bai-Zhong Zhang, Hong-Liang Wang
Pestic Biochem Physiol. doi: 10.1016/j.pestbp.2022.105133.
83. Zhiyuan Niu, Zhixia Luo, Pengyang Sun, Linwei Ning, Xinru Jin, Guanxu Chen, Changjiang Guo, Lingtong Zhi, Wei Chang, Wuling Zhu
Biodes Res. doi: 10.34133/2022/9823578.
84. Shaojin Li, Yaozhong Liang, Jianyu Zou, Zhenbin Cai, Hua Yang, Jie Yang, Yunlong Zhang, Hongsheng Lin, Guowei Zhang, Minghui Tan
J Biol Chem. doi: 10.1016/j.jbc.2022.102292. (IF 4.238)
85. Bai-Zhong Zhang, Meng-Yuan Zhang, Ya-She Li, Gui-Lei Hu, Xin-Zheng Fan, Tian-Xin Guo, Feng Zhou, Pei Zhang, Yan-Bing Wu, Yang-Fan Gao, Xi-Wu Gao
Pestic Biochem Physiol. doi: 10.1016/j.pestbp.2022.105218.
86. Gui-Lei Hu, Liu-Yang Lu, Ya-She Li, Xu Su, Wen-Yang Dong, Bai-Zhong Zhang, Run-Qiang Liu, Ming-Wang Shi, Hong-Liang Wang, Xi-Ling Chen
Bull Entomol Res. doi: 10.1017/S0007485322000037.
87. Haibo Fang, Xiu Wang, Xiaoxia Liu, J P Michaud, Yanan Wu, Huaijiang Zhang, Yisong Li, Zhen Li
Insect Mol Biol. doi: 10.1111/imb.12794. (IF 2.533)