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试剂> 常用试剂> 抗生素与防腐剂> 哺乳动物细胞培养

Puromycin Dihydrochloride (嘌呤霉素)

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产品编号: ST551-10mg

产品包装:10mg/ml×1ml

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10mg/ml×1ml 10mg/ml×5ml 250mg

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产品编号	产品名称	产品包装	产品价格
ST551-10mg	Puromycin Dihydrochloride (嘌呤霉素)	10mg/ml×1ml	208.00元
ST551-50mg	Puromycin Dihydrochloride (嘌呤霉素)	10mg/ml×5ml	868.00元
ST551-250mg	Puromycin Dihydrochloride (嘌呤霉素)	250mg	3298.00元

      Puromycin是来源于Streptomyces alboniger的一种氨基核苷类抗生素，中文名为嘌呤霉素，常用于筛选通过质粒转染/转
化、病毒感染等方法能表达pac基因(puro^r)的真核或原核多克隆或单克隆细胞。Puromycin不仅用于稳定细胞株的筛选，也用于稳定细胞株的维持。本产品的10mg/ml包装经过滤除菌，可以直接用于细胞培养。
      Puromycin的特点是快速作用于细胞，一般2天内可以杀死99%的不表达pac基因的细胞。
      在革兰氏阳性菌、动物或昆虫细胞中，嘌呤霉素通过抑制蛋白质合成而抑制或杀死细胞。其作用机制为嘌呤霉素是氨酰-tRNA分子3'末端的类似物，能够与核糖体的A位点结合并掺入到延伸的肽链中。嘌呤霉素与A位点结合后，不会参与随后的任何反应，从而导致蛋白质合成的提前终止并释放出C-末端含有嘌呤霉素的不成熟多肽。
      Pac基因表达嘌呤霉素N-乙酰转移酶(Puromycin N-acetyl-tranferase)，该基因是在Streptomyces alboniger中发现的。如果表达基因，就会对嘌呤霉素产生抗性，这一特性目前普遍应用于筛选表达pac基因的哺乳动物稳定细胞株等。例如，很多商业化的慢病毒载体都携带pac基因(一般在质粒图谱上标记为puro^r)，从而利用嘌呤霉素筛选特定基因的稳定表达细胞株。嘌呤霉素也可以用来筛选表达pac基因的大肠杆菌菌株、酵母菌株等。
      化学性质

化学名	(2S)-2-amino-N-[(2S,3S,4R,5R)-5-[6-(dimethylamino)purin-9-yl]-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl]-3-(4-methoxyphenyl)propanamide;dihydrochloride
化学式	C₂₂H₂₉N₇O₅•2HCl
分子量	544.43
CAS号	58-58-2
纯度	>98%

本产品10mg/ml包装配制在20mM HEPES(pH7.4)溶液中，浓度为10mg/ml，经0.22μm滤膜过滤除菌，可以直接用于细胞培养。本产品250mg包装为粉末装。
包装清单：

产品编号	产品名称	包装
ST551-10mg	Puromycin Dihydrochloride (嘌呤霉素)	10mg/ml×1ml
ST551-50mg	Puromycin Dihydrochloride (嘌呤霉素)	10mg/ml×5ml
ST551-250mg	Puromycin Dihydrochloride (嘌呤霉素)	250mg
—	说明书	1份

保存条件：
        -20℃保存，至少一年有效。10mg/ml包装避免反复冻融。
注意事项：
       本产品对人体有害，操作时请小心，并注意有效防护以避免直接接触人体或吸入体内。
       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。

1. 推荐工作浓度：推荐的作用于哺乳动物细胞的嘌呤霉素浓度一般为1-10μg/ml，但最佳工作浓度需要通过剂量反应曲线来确定。
表1. 部分常见哺乳动物细胞的推荐工作浓度表

细胞名称	细胞类型	嘌呤霉素浓度
A549	Lung cancer	1-2μg/ml
B16	Mouse melanocytes	1-2μg/ml
ES cell	Human embryonic stem cells	0.5-5μg/ml
H1299	Non-small cell lung carcinoma	1-3μg/ml
HEK293	Human embryonic kidney	0.5-3μg/ml
HeLa	Human cervical cancer	1-2μg/ml
HepG2	Human hepatocellular carcinoma	0.5-2μg/ml
HT1080	Human fibrosarcoma	0.5-2μg/ml
MCF-7	Human breast cancer	0.5-2μg/ml
MDA-MB-231	Human breast cancer	0.5-5μg/ml
MEF	Mouse fibroblasts	1-5μg/ml

2. 嘌呤霉素剂量反应曲线的确定(以shRNA转染或者慢病毒感染为例）
嘌呤霉素的有效筛选浓度跟细胞类型、生长状态、细胞密度、细胞代谢及细胞所处细胞周期等因素相关。为了筛选到稳定表达的shRNA或感染病毒的细胞株，确定杀死未转染/感染细胞的最低浓度嘌呤霉素非常重要。对于初次使用的细胞，一般需要通过实验来确定适合自身实验体系的剂量反应曲线(dose-response curve or kill curve)。
a. 第一天：24孔板中以5~8×10⁴cells/孔的密度接种细胞，接种够量的孔以便进行后续的剂量梯度实验。细胞培养箱内培养过夜。
b. 第二天：在培养过夜后的细胞中更换新鲜配制的筛选培养基，该筛选培养基为含不同浓度嘌呤霉素的新鲜培养基(如0、1、2.5、5、7.5、10μg/ml等)，更换培养基后在细胞培养箱中继续培养。
c. 第三天后：由于嘌呤霉素可以快速作用于细胞，一般2天内可以杀死99%的未表达pac基因的细胞，所以在加嘌呤霉素后的1-2天就可以进行观察细胞存活率，从而确定有效杀死正常细胞的药物最低浓度。如果细胞耐药性比较强，需要每日观察，一般4-10天内即可确定嘌呤霉素的最低浓度。
3. 哺乳动物稳定表达细胞株的筛选
转染含有pac基因的质粒或者感染含有该基因的病毒后，即可筛选稳定表达株。
a. 细胞转染或感染48小时后，将细胞置于含有适当浓度嘌呤霉素的新鲜培养基中培养，此为处理组。
注意：当细胞处于分裂活跃期时，抗生素作用最明显。细胞过于密集，抗生素产生的效力会明显下降，所以细胞的密度最好不超过25%。建议同时做一个正常细胞的对照组。转染或感染48小时后，如果细胞过密也可以消化后重新接种细胞，培养过夜后即可进行嘌呤霉素筛选。
b. 每隔2-3天，更换含有嘌呤霉素的培养基。
c. 筛选7天后，对照组正常细胞应该100%死亡，处理组中存活的细胞为表达pac基因的细胞。然后根据实验目的进行多克隆或单克隆细胞的筛选。
注意：每日进行细胞生长状态的观察。嘌呤霉素的筛选至少需要24小时，有效浓度嘌呤霉素的筛选周期一般在2-10天。
d. 待细胞可以稳定生长后，嘌呤霉素的浓度可以减半用于后续的培养。在得到稳定表达细胞株后，一般建议嘌呤霉素也须持续加入，并2-3天更换含有嘌呤霉素的新鲜培养液。

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42. Lihui Lu, Jianwei Wang, Fang Fang, Ailian Guo, Shuting Jiang, Yanfang Tao, Yongping Zhang, Yan Li, Kunlong Zhang, Zimu Zhang, Ran Zhuo, Xinran Chu, Xiaolu Li, Yuanyuan Tian, Li Ma, Xu Sang, Yanling Chen, Juanjuan Yu, Yang Yang, Haibo Cao, Jizhao Gao, Jun Lu, Shaoyan Hu, Jian Pan, Hailong He

LMO2 promotes the development of AML through interaction with transcription co-regulator LDB1

Cell Death Dis. 2023 Aug 12;14(8):518. doi: 10.1038/s41419-023-06039-w.

43. Zhihuan You, Xiaotian Ye, Meihua Jiang, Ning Gu, Caihong Liang

lnc-MRGPRF-6:1 Promotes ox-LDL-Induced Macrophage Ferroptosis via Suppressing GPX4

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44. Xiangyu Wang, Minkai Song, Xiaomin Li, Cailin Su, Yanlin Yang, Kai Wang, Cuiting Liu, Zongji Zheng, Yijie Jia, Shijing Ren, Wenhui Dong, Jiaqi Chen, Ting Wang, Lerong Liu, Meiping Guan, Chao Zhang, Yaoming Xue

CERS6-derived ceramides aggravate kidney fibrosis by inhibiting PINK1-mediated mitophagy in diabetic kidney disease

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45. Xiangxiang Liu, Jian Qin, Junjie Nie, Rui Gao, Shangshang Hu, Huiling Sun, Shukui Wang, Yuqin Pan

ANGPTL2+cancer-associated fibroblasts and SPP1+macrophages are metastasis accelerators of colorectal cancer

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46. Ling Zhang, Jun Zhang, Xiaofeng Xuan, Di Wu, Jianfeng Yu, Peizhen Wang, Xiaomei Yang, Jieru Zhang, Wenjuan Gan, Mengfan He, Xiao-Min Liu, Jun Zhou, Donglai Wang, Wei Gu, Dawei Li

A p53/LINC00324 positive feedback loop suppresses tumor growth by counteracting SET-mediated transcriptional repression

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47. Biying Yang, Jingrui Pan, Xiao-Ni Zhang, Hongxuan Wang, Lei He, Xiaoming Rong, Xiangpen Li, Ying Peng

NRF2 activation suppresses motor neuron ferroptosis induced by the SOD1G93A mutation and exerts neuroprotection in amyotrophic lateral sclerosis

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48. Yue Xiao, Xinyi Chen, Weiwei Hu, Wenjing Ma, Qianqian Di, Haimei Tang, Xibao Zhao, Guodong Huang, Weilin Chen

USP39-mediated deubiquitination of Cyclin B1 promotes tumor cell proliferation and glioma progression

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49. Kailong Ji, Wei Liu, Weihang Yin, Xiangrong Kong, Honghong Xu, Zeng-Wei Lai, Jing-Ya Li, Jian-Min Yue

A new class of potent liver injury protective compounds: Structural elucidation, total synthesis and bioactivity study

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50. Yuanyi Wang, Ting Zhang, Hongfei Du, Min Yang, Guangsu Xie, Teng Liu, Shihua Deng, Wei Yuan, Shuang He, Dongming Wu, Ying Xu

Dipeptidase‑2 is a prognostic marker in lung adenocarcinoma that is correlated with its sensitivity to cisplatin

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51. Tingting Feng, Ru Zhao, Hanwen Zhang, Feifei Sun, Jing Hu, Meng Wang, Mei Qi, Ling Liu, Lin Gao, Yabo Xiao, Junhui Zhen, Weiwen Chen, Lin Wang, Bo Han

Reciprocal negative feedback regulation of ATF6α and PTEN promotes prostate cancer progression

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52. Rongfeng Huang, Jianghui Chen, Meiyu Zhou, Haoran Xin, Sin Man Lam, Xiaoqing Jiang, Jie Li, Fang Deng, Guanghou Shui, Zhihui Zhang, Min-Dian Li

Multi-omics profiling reveals rhythmic liver function shaped by meal timing

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53. Yanli Zhang, Yulin Zhang, Ye Lei, Junrong Wu, Yiyuan Kang, Shuo Zheng, Longquan Shao

MDM2 upregulation induces mitophagy deficiency via Mic60 ubiquitination in fetal microglial inflammation and consequently neuronal DNA damage caused by exposure to ZnO-NPs during pregnancy

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54. Xiaojun Dai, Yanwei Yu, Chen Zou, Bo Pan, Haibo Wang, Shanshan Wang, Xiaojuan Wang, Chenghai Wang, Dongmei Liu, Yanqing Liu

Traditional Banxia Xiexin decoction inhibits invasion, metastasis, and epithelial mesenchymal transition in gastric cancer by reducing lncRNA TUC338 expression

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55. Shenghua Zhang, Zheng Zhang, Xiaolong Liu, Yibin Deng, Jian Zheng, Jieqiong Deng, Yirong Wang, Binbin Guo, Fanrong Li, Xiaoyue Chen, Yacheng Pan, Jieyu Wang, Jiachun Lu, Siqi Wu, Qiang Guo, Yifeng Zhou

LncRNA-Encoded Micropeptide ACLY-BP Drives Lipid Deposition and Cell Proliferation in Clear Cell Renal Cell Carcinoma via Maintenance of ACLY Acetylation

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56. Meijian Liao, Xiaolin Sun, Wendan Zheng, Mengdi Wu, Yifan Wang, Jia Yao, Yu Ma, Shoucui Gao, Dongsheng Pei

LINC00922 decoys SIRT3 to facilitate the metastasis of colorectal cancer through up-regulation the H3K27 crotonylation of ETS1 promoter

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57. Zhou Yang, Xiyi Wei, Chengjian Ji, Xiaohan Ren, Wei Su, Yichun Wang, Jingwan Zhou, Zheng Zhao, Pengcheng Zhou, Kejie Zhao, Bing Yao, Ninghong Song, Chao Qin

OGT/HIF-2α axis promotes the progression of clear cell renal cell carcinoma and regulates its sensitivity to ferroptosis

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58. Wen Tian, Jinhui Zhao, Wenzhong Wang

Targeting CDH17 with Chimeric Antigen Receptor-Redirected T Cells in Small Cell Lung Cancer

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59. Zhen Li, Kang Ning, Diwei Zhao, Zhaohui Zhou, Junliang Zhao, Xingbo Long, Zhenyu Yang, Dong Chen, XinYang Cai, Lexuan Hong, Luyao Zhang, Fangjian Zhou, Jun Wang, Yonghong Li

Targeting the Metabolic Enzyme PGAM2 Overcomes Enzalutamide Resistance in Castration-Resistant Prostate Cancer by Inhibiting BCL2 Signaling

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60. Yuanyuan Liu, Xing Wang, Yuying Liu, Jianqiang Yang, Wei Mao, Chen Feng, Xiaoliang Wu, Xinwei Chen, Lixiao Chen, Pin Dong

N4-acetylcytidine-dependent GLMP mRNA stabilization by NAT10 promotes head and neck squamous cell carcinoma metastasis and remodels tumor microenvironment through MAPK/ERK signaling pathway

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61. Minghui Deng, Yefeng Xu, Yongwei Yao, Yiqing Wang, Qingying Yan, Miao Cheng, YunXia Liu

Circular RNA hsa_circ_0051246 acts as a microRNA-375 sponge to promote the progression of gastric cancer stem cells via YAP1

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Baicalein improves the chemoresistance of ovarian cancer through regulation of CirSLC7A6

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Annexin A5 derived from matrix vesicles protects against osteoporotic bone loss via mineralization

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64. Xiaoxiong Zou, Yu Xie, Zhongfei Zhang, Zhiming Feng, Jianbang Han, Qian Ouyang, Shiting Hua, Sixian Huang, Cong Li, Zhizheng Liu, Yingqian Cai, Yuxi Zou, Yanping Tang, Haijia Chen, Xiaodan Jiang

MCPIP-1 knockdown enhances endothelial colony-forming cell angiogenesis via the TFRC/AKT/mTOR signaling pathway in the ischemic penumbra of MCAO mice

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65. Song Wang, Jing Ma, Xin Li, Xinmiao Xian, Guikun Tan, Hongwei Cai, Bingwu Yang, Anqi Zhang, Jianran Guo, Guohao Gu, Zhen Meng, Bo Fu

EGR-1 Contributes to Pulmonary Edema by Regulating the Epithelial Sodium Channel in Lipopolysaccharide-Induced Acute Lung Injury

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66. Ling Zhang, Mengfan He, Peizhen Wang, Jianfeng Yu, Dawei Li

A protocol to investigate the effects of lncRNAs on in vivo protein-protein interactions using proximity ligation assay

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67. Xiangfei Sun, Qiang Zhang, Xiaohan Lin, Ping Shu, Xiaodong Gao, Kuntang Shen

Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination

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68. Feiyu Tang, Can Lu, Xiang He, Wei Lin, Bowen Xie, Xing Gao, Yang Peng, Desong Yang, Lunquan Sun, Liang Weng

E3 ligase Trim35 inhibits LSD1 demethylase activity through K63-linked ubiquitination and enhances anti-tumor immunity in NSCLC

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69. Shuo Yang, Ping Zhou, Lelei Zhang, Xiangpeng Xie, Yuanyi Zhang, Kaida Bo, Jing Xue, Wei Zhang, Faxue Liao, Pengfei Xu, Yong Hu, Ruyu Yan, Dan Liu, Jun Chang, Kecheng Zhou

VAMP8 suppresses the metastasis via DDX5/β-catenin signal pathway in osteosarcoma

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70. Li Xiong, Yingying Gong, Huashan Liu, Liang Huang, Ziwei Zeng, Xiaobin Zheng, Wenxin Li, Zhenxing Liang, Liang Kang

circGlis3 promotes β-cell dysfunction by binding to heterogeneous nuclear ribonucleoprotein F and encoding Glis3-348aa protein

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71. Chuanlai Yang, Yuegang Ge, Yachen Zang, Ming Xu, Lu Jin, Yang Wang, Xinyu Xu, Boxin Xue, Zhiwei Wang, Lixia Wang

CDC20 promotes radioresistance of prostate cancer by activating Twist1 expression

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Host factor MxA restricts Dabie bandavirus infection by targeting the viral NP protein to inhibit NP-RdRp interaction and ribonucleoprotein activity

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73. Xiaozhou Xie, Yulong Zhao, Fengying Du, Baoshan Cai, Zhen Fang, Yuan Liu, Yaodong Sang, Chenghao Ma, Zhaodong Liu, Xinshuai Yu, Chi Zhang, Jiayu Jiang, Zi Gao, Yan Liu, Xiaoyan Lin, Haiyan Jing, Xiuming Zhong, Lei Cong, Honghai Dai, Dan Sha, Na Shao, Hong Feng, Leping Li, Jin Liu, Liang Shang

Pan-cancer analysis of the tumorigenic role of Fanconi anemia complementation group D2 (FANCD2) in human tumors

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74. Xue Yang, Chun Gu, Jingshu Cai, Feiyang Li, Xing He, Lunan Luo, Wengan Xiao, Boyan Hu, Jing Hu, Hao Qian, Shangqing Ren, Lin Zhang, Xianjun Zhu, Lu Yang, Jing Yang, Zhenglin Yang, Yi Zheng, Xiang Huang, Ziyan Wang

Excessive SOX8 reprograms energy and iron metabolism to prime hepatocellular carcinoma for ferroptosis

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75. Juanjuan Ji, Zhi Zhou, Qi Luo, Yaling Zhu, Rui Wang, Yali Liu

TMEM16A enhances the activity of the Cdc42-NWASP signaling pathway to promote invasion and metastasis in oral squamous cell carcinoma

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76. Peng-Cheng Chen, Yong Ning, Hui Li, Jin-Gen Su, Jiang-Bo Shen, Qing-Chun Feng, Shu-Heng Jiang, Pei-Dong Shi, Run-Sheng Guo

Targeting ONECUT3 blocks glycolytic metabolism and potentiates anti-PD-1 therapy in pancreatic cancer

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77. Qinghua Zeng, Jingjing Cao, Fei Xie, Lina Zhu, Xiangdong Wu, Xifeng Hu, Zheng Chen, Xiaoqing Chen, Xiangzhi Li, Cheng-Ming Chiang, Huansheng Wu

CRISPR-Cas9-mediated chicken prmt5 gene knockout and its critical role in interferon regulation

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78. Jingyue Yang, Yindi Xiong, Herman W Barkema, Xiaofang Tong, Yushan Lin, Zhaoju Deng, John P Kastelic, Diego B Nobrega, Yue Wang, Bo Han, Jian Gao