萤火虫萤光素酶报告基因检测试剂盒(RG006)

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萤火虫萤光素酶报告基因检测试剂盒

产品编号: RG006

产品包装:1000次
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2316.00 10

价格: ¥ 2316.00

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产品编号 产品名称 产品包装 产品价格
RG005 萤火虫萤光素酶报告基因检测试剂盒 100次 412.00元
RG006 萤火虫萤光素酶报告基因检测试剂盒 1000次 2316.00元

碧云天生产的萤火虫萤光素酶报告基因检测试剂盒(Firefly Luciferase Reporter Gene Assay Kit)，是一种以萤光素(luciferin)为底物来检测萤火虫萤光素酶(firefly luciferase)活性的试剂盒。
萤火虫萤光素酶是一种分子量约为61kD的蛋白，在ATP、镁离子和氧气存在的条件下，可以催化luciferin氧化成oxyluciferin。在luciferin氧化的过程中，会发出生物萤光(bioluminescence)。生物萤光可以通过化学发光仪(luminometer)或液闪测定仪进行测定。。本试剂盒的检测原理参考图1。

萤火虫萤光素酶报告基因检测试剂盒(RG006)
图1. 萤火虫萤光素酶的检测原理图。

通过萤光素和萤光素酶这一生物发光体系，可以非常灵敏、高效地检测基因的表达。通常把感兴趣 基因的转录调控元件或5’启动子区克隆在luciferase的上游，或把3’-UTR区克隆在luciferase的下游等，构建成报告基因(reporter gene)质粒。然后转染细胞，用适当药物等处理细胞后裂解细胞，测定萤光素酶活性。通过萤光素酶活性的高低来判断药物处理等对目的基因的转录调控作用。
萤火虫萤光素酶催化luciferin发光的最强发光波长为560nm(centered around 560nm)。
本试剂盒可以测定1000个样品。
包装清单：

产品编号 产品名称 包装
RG006-1 报告基因细胞裂解液 RG005-1×10
RG006-2 萤火虫萤光素酶检测试剂 RG005-2×10
— 说明书 1份

保存条件：
报告基因细胞裂解液4℃保存3个月有效，-20℃保存一年有效；萤光素酶检测试剂-20℃避光保存6个月有效，-80℃避光保存一年有效。
注意事项：
为取得最佳测定效果，在用单管的化学发光仪测定时，样品和测定试剂混合后到测定前的时间应尽
量控制在相同时间内，例如30秒内；使用具有化学发光测定功能的多功能荧光酶标仪时，宜先把样品全部加好，然后统一加入萤火虫萤光素酶检测试剂。
由于温度对酶反应有影响，所以测定时样品和试剂均需达到室温后再进行测定。
为保证萤光素酶检测试剂的稳定性，可以采取适当分装后避光保存的方法，以避免反复冻融和长时间暴露于室温。经测试，反复冻融三次，对测定结果无明显影响。
样品和测定试剂混合后，必须等待1-2秒，再进行测定。测定时间通常为10秒，根据情况也可以测定更长或更短时间，但是同一批样品宜使用相同的测定时间。
为避免由于质粒转细胞时效率的差异而带来的误差，可以同时转入海肾萤光素酶(Renilla luciferase)的报告基因质粒作为内参，采用碧云天的双萤光素酶报告基因检测试剂盒(RG027/RG028)进行检测；也可以同时转入β-半乳糖苷酶(β-galactosidase,β-gal)报告基因质粒作为内参，然后采用碧云天生产的β-半乳糖苷酶报告基因检测试剂盒(RG0036)进行检测。采用本试剂盒中的报告基因细胞裂解液裂解获得的样品可以直接用于β-半乳糖苷酶报告基因检测试剂盒(RG0036)的检测。
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. 裂解细胞：将报告基因细胞裂解液充分混匀后，按如下方式加入报告基因细胞裂解液，充分裂解细胞。
a. 对于贴壁细胞：吸尽细胞培养液后，参考下表加入适量的报告基因细胞裂解液；对于悬浮细胞：离心去上清后，参考下表加入适量报告基因细胞裂解液。

器皿类型 96孔板 48孔板 24孔板 12孔板 6孔板
报告基因细胞裂解液 (微升/孔) 100 150 200 300 500

注：如果荧光素酶的表达水平比较低，可以尝试使用更少的裂解液，例如6孔板的每孔用量可以最小为100微升。
b. 充分裂解后，10,000-15,000g离心3-5分钟，取上清用于测定。
注：细胞裂解后可立即测定萤光素酶，也可以先冻存，待以后再测定。冻存样品需融解，并达到室温后再进行测定。
2. 融解萤光素酶检测试剂，并达到室温。
3. 按仪器操作说明书开启化学发光仪或具有检测化学发光功能的多功能酶标仪，将测定间隔设为2秒，测定时间设为10秒。
4. 每个样品测定时，取样品20-100微升(如果样品量足够，请加入100微升；如果样品量不足可以适当减少用量，但同批样品的使用量宜保持一致)。
5. 加入100微升萤光素酶检测试剂，用枪打匀或用其它适当方式混匀后测定RLU(relative lightunit)。以报告基因细胞裂解液为空白对照。本试剂盒的检测效果以及与同类竞争产品的检测效果比较可以参考图2。

萤火虫萤光素酶报告基因检测试剂盒(RG006)

图2. 本产品和P公司竞争产品(Competitor P)对相同样品的检测效果。实际读数会因细胞数量、转染效果、检测仪器等的不同而存在差异，图中数据仅供参考。

常见问题：
1. Luminometer和荧光分光光度计有何不同？
荧光分光光度计检测的样品本身不能发光，样品需要由特定波长的激发光激发，然后才能产生荧光并被荧光分光光度计检测。Luminometer检测的样品本身可以发光，不需要激发光进行激发。也就是说luminometer是检测化学发光(萤光)的仪器。有些型号的荧光分光光度计也具有luminometer的功能，即也可以检测化学发光。您所使用的荧光分光光度计能否用于化学发光的测定请仔细阅读该仪器的说明书。
2. 可以进行ATP化学发光检测的仪器是否就可以用于本试剂盒的检测？
是。ATP化学发光的检测原理和本试剂盒的原理相同，可以用相同的仪器测定。

相关产品：

产品编号 产品名称 包装
RG005 萤火虫萤光素酶报告基因检测试剂盒 100次
RG006 萤火虫萤光素酶报告基因检测试剂盒 1000次
RG016 海肾萤光素酶报告基因检测试剂盒 100次
RG017 海肾萤光素酶报告基因检测试剂盒 1000次
RG027 双萤光素酶报告基因检测试剂盒 100次
RG028 双萤光素酶报告基因检测试剂盒 1000次
RG0036 β-半乳糖苷酶报告基因检测试剂盒 200次

萤火虫萤光素酶报告基因检测试剂盒(RG006)

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56. Chao Yang,Zeqiang Yan,Fen Hu,Wei Wei,Zhihua Sun,Wei Xu
Silencing of microRNA-517a induces oxidative stress injury in melanoma cells via inactivation of the JNK signaling pathway by upregulating CDKN1C
Cancer Cell Int. 2020 Jan 29;20:32.;doi: 10.1186/s12935-019-1064-y. (IF 4.175)

57. Li Jiang,Yanguo Qiao,Zhenghui Wang,Xiuzhu Ma,Haichao Wang,Jian Li
Inhibition of microRNA-103 attenuates inflammation and endoplasmic reticulum stress in atherosclerosis through disrupting the PTEN-mediated MAPK signaling
J Cell Physiol. 2020 Jan;235(1):380-393.;doi: 10.1002/jcp.28979. (IF 5.546)

58. Xiao-Lu Guan,Bao-Cun Zhang,Li Sun
Japanese flounder pol-miR-3p-2 suppresses Edwardsiella tarda infection by regulation of autophagy via p53
Dev Comp Immunol. 2020 Feb;103:103531.;doi: 10.1016/j.dci.2019.103531. (IF 3.192)

59. Jiwei Li,Li Wei,Zhijun Han,Zhong Chen,Quan Zhang
Long non-coding RNA X-inactive specific transcript silencing ameliorates primary graft dysfunction following lung transplantation through microRNA-21-dependent mechanism
EBioMedicine. 2020 Feb;52:102600.;doi: 10.1016/j.ebiom.2019.102600. (IF 5.736)

60. Li Chen,De-Zhong Sun,Yu-Gui Fu,Pei-Zhen Yang,Huai-Qing Lv,Yongli Gao,Xiao-Yan Zhang
Upregulation of microRNA-141 suppresses epithelial-mesenchymal transition and lymph node metastasis in laryngeal cancer through HOXC6-dependent TGF-β signaling pathway
Cell Signal. 2020 Feb;66:109444.;doi: 10.1016/j.cellsig.2019.109444. (IF 3.968)

61. Wenfeng Xie,Lei Chen,Li Chen,Qiuye Kou
Silencing of long non-coding RNA MALAT1 suppresses inflammation in septic mice: role of microRNA-23a in the down-regulation of MCEMP1 expression
Inflamm Res. 2020 Feb;69(2):179-190.;doi: 10.1007/s00011-019-01306-z. (IF 3.174)

62. Ruijuan Qi,Ximeng Li,Xiaoyu Zhang,Yunfeng Huang,Qiaoling Fei,Yixin Han,Runlan Cai,Yuan Gao,Yun Qi
Ethanol extract of Elephantopus scaber Linn. Attenuates inflammatory response via the inhibition of NF-κB signaling by dampening p65-DNA binding activity in lipopolysaccharide-activated macrophages
J Ethnopharmacol. 2020 Mar 25;250:112499.;doi: 10.1016/j.jep.2019.112499. (IF 3.69)

63. Peng Xia,Rui Gu,Wei Zhang,Yi-Fu Sun
lncRNA CEBPA-AS1 Overexpression Inhibits Proliferation and Migration and Stimulates Apoptosis of OS Cells via Notch Signaling
MOL THER-NUCL ACIDS. 2020 Mar 6;19:1470-1481.;doi: 10.1016/j.omtn.2019.10.017. (IF 7.032)

64. Duo-Ping Wang,Xiao-Zhun Tang,Quan-Kun Liang,Xian-Jie Zeng,Jian-Bo Yang,Jian Xu
microRNA-599 promotes apoptosis and represses proliferation and epithelial-mesenchymal transition of papillary thyroid carcinoma cells via downregulation of Hey2-depentent Notch signaling pathway
J Cell Physiol. 2020 Mar;235(3):2492-2505.;doi: 10.1002/jcp.29154. (IF 5.546)

65. Jing Ma,Zhijian Zhou
Downregulation of miR-302b is associated with poor prognosis and tumor progression of breast cancer
Breast Cancer. 2020 Mar;27(2):291-298.;doi: 10.1007/s12282-019-01022-w. (IF 2.695)

66. Shuang Liu,Xian-Hui Ning,Xiao-Lu Guan,Xue-Peng Li,Li Sun
Characterization of Streptococcus iniae-induced microRNA profiles in Paralichthys olivaceus and identification of pol-3p-10740_175 as a regulator of antibacterial immune response
FISH SHELLFISH IMMUN. 2020 Mar;98:860-867.;doi: 10.1016/j.fsi.2019.11.045. (IF 3.298)

67. Jinzhang Cheng,Junjun Chen,Yin Zhao,Jingpu Yang,Kai Xue,Zonggui Wang
MicroRNA-761 suppresses remodeling of nasal mucosa and epithelial-mesenchymal transition in mice with chronic rhinosinusitis through LCN2
Stem Cell Res Ther. 2020 Apr 9;11(1):151.;doi: 10.1186/s13287-020-01598-7. (IF 5.116)

68. Lei Jia,Yuwen Song,Luyan Mu,Qingla Li,Jiabin Tang,Zhao Yang,Wenjuan Meng
Long noncoding RNA TPT1-AS1 downregulates the microRNA-770-5p expression to inhibit glioma cell autophagy and promote proliferation through STMN1 upregulation
J Cell Physiol. 2020 Apr;235(4):3679-3689.;doi: 10.1002/jcp.29262. (IF 5.546)

69. Lilei Peng,Yang Ming,Ling Zhang,Jie Zhou,Wei Xiang,Shan Zeng,Haiping He,Ligang Chen
MicroRNA-30a suppresses self-renewal and tumorigenicity of glioma stem cells by blocking the NT5E-dependent Akt signaling pathway
FASEB J. 2020 Apr;34(4):5128-5143.;doi: 10.1096/fj.201802629RR. (IF 4.966)

70. Jinqiu Li,Xueshibojie Liu,Shanji Nan,Chengbi Xu
Silencing of long non-coding RNA LINC00520 promotes radiosensitivity of head and neck squamous cell carcinoma cells
FREE RADICAL RES. 2020 Apr;54(4):254-270.;doi: 10.1080/10715762.2020.1752373. (IF 2.839)

71. Wei Qian,Fengbo Jin,Yiming Zhao,Yingying Chen,Ling Ge,Lixia Liu,Mingzhen Yang
Downregulation of microRNA-144 inhibits proliferation and promotes the apoptosis of myelodysplastic syndrome cells through the activation of the AKAP12-dependent ERK1/2 signaling pathway
Cell Signal. 2020 Apr;68:109493.;doi: 10.1016/j.cellsig.2019.109493. (IF 3.968)

72. Na Han,Hai-Yan Fang,Jie-Xuan Jiang,Qian Xu
Downregulation of microRNA-873 attenuates insulin resistance and myocardial injury in rats with gestational diabetes mellitus by upregulating IGFBP2
AM J PHYSIOL-ENDOC M. 2020 May 1;318(5):E723-E735.;doi: 10.1152/ajpendo.00555.2018. (IF 3.469)

73. Qing Meng,Bing Qiu
Exosomal MicroRNA-320a Derived From Mesenchymal Stem Cells Regulates Rheumatoid Arthritis Fibroblast-Like Synoviocyte Activation by Suppressing CXCL9 Expression
Front Physiol. 2020 May 26;11:441.;doi: 10.3389/fphys.2020.00441. (IF 3.367)

74. Wen-Rui Li,Xiao-Lu Guan,Shuai Jiang,Li Sun
The novel fish miRNA pol-miR-novel_171 and its target gene FAM49B play a critical role in apoptosis and bacterial infection
Dev Comp Immunol. 2020 May;106:103616.;doi: 10.1016/j.dci.2020.103616. (IF 3.192)

75. Jie Chen,Mao Liu,Xiao Luo,Lihui Peng,Zixia Zhao,Chengsong He,Yue He
Exosomal miRNA-486-5p derived from rheumatoid arthritis fibroblast-like synoviocytes induces osteoblast differentiation through the Tob1/BMP/Smad pathway
BIOMATER SCI-UK. 2020 Jun 21;8(12):3430-3442.;doi: 10.1039/c9bm01761e. (IF 6.183)

76. Chunyan Li,Tingfeng Han,Run Li,Liming Fu,Lei Yue
miR-26a-5p mediates TLR signaling pathway by targeting CTGF in LPS-induced alveolar macrophage
BIOSCIENCE REP. 2020 Jun 26;40(6):BSR20192598.;doi: 10.1042/BSR20192598. (IF 2.942)

77. Junling Gong,Haiying Fan,Jing Deng,Qiumei Zhang
LncRNA HAND2-AS1 represses cervical cancer progression by interaction with transcription factor E2F4 at the promoter of C16orf74
J Cell Mol Med. 2020 Jun;24(11):6015-6027.;doi: 10.1111/jcmm.15117. (IF 4.486)

78. Xiao Gu,Xiaocui Yao,Dengtao Liu
Up-regulation of microRNA-335-5p reduces inflammation via negative regulation of the TPX2-mediated AKT/GSK3β signaling pathway in a chronic rhinosinusitis mouse model
Cell Signal. 2020 Jun;70:109596.;doi: 10.1016/j.cellsig.2020.109596. (IF 3.968)

79. Wenchao Zhong,Xingyang Li,Janak L Pathak,Liangjiao Chen,Wei Cao,Mingjing Zhu,Qianting Luo,Antong Wu,Yunxin Chen,Lingbo Yi,Manyuan Ma,Qingbin Zhang
Dicalcium silicate microparticles modulate the differential expression of circRNAs and mRNAs in BMSCs and promote osteogenesis via circ_1983-miR-6931-Gas7 interaction
BIOMATER SCI-UK. 2020 Jul 7;8(13):3664-3677.;doi: 10.1039/d0bm00459f. (IF 6.183)

80. Xiaohe Fang,Yong Dong,Ruilin Yang,Lunshou Wei
LINC00619 restricts gastric cancer progression by preventing microRNA-224-5p-mediated inhibition of OPCML
Arch Biochem Biophys. 2020 Aug 15;689:108390.;doi: 10.1016/j.abb.2020.108390. (IF 3.391)

81. Wenbiao Liao,Yi Zhang
MicroRNA-381 facilitates autophagy and apoptosis in prostate cancer cells via inhibiting the RELN-mediated PI3K/AKT/mTOR signaling pathway
Life Sci. 2020 Aug 1;254:117672.;doi: 10.1016/j.lfs.2020.117672. (IF 3.647)

82. Yuedi Ding,Jun Fan,Lili Deng,Ying Peng,Bin Zhou,Biao Huang
Evaluation of Tumor Specificity and Immunity of Thymidine Kinase-Deleted Vaccinia Virus Guang9 Strain
ONCOTARGETS THER. 2020 Aug 5;13:7683-7697.;doi: 10.2147/OTT.S260288. (IF 3.337)

83. Zhuo Liang,Yue Luo,Yonggang Lv
Mesenchymal stem cell-derived microvesicles mediate BMP2 gene delivery and enhance bone regeneration
J Mater Chem B. 2020 Aug 5;8(30):6378-6389.;doi: 10.1039/d0tb00422g. (IF 5.344)

84. Guo-Feng Zhang,Jia-Cheng Wu,Hong-Yong Wang,Wei-Dong Jiang,Ling Qiu
Overexpression of microRNA-205-5p exerts suppressive effects on stem cell drug resistance in gallbladder cancer by down-regulating PRKCE
BIOSCIENCE REP. 2020 Sep 30;40(9):BSR20194509.;doi: 10.1042/BSR20194509. (IF 2.942)

85. Hui Zhao,Feng Ding,Guanghong Zheng
LncRNA TMPO-AS1 promotes LCN2 transcriptional activity and exerts oncogenic functions in ovarian cancer
FASEB J. 2020 Sep;34(9):11382-11394.;doi: 10.1096/fj.201902683R. (IF 4.966)

86. Zhiyan Ruan,Hongling Deng,Minhua Liang,Zhe Xu,Manxiang Lai,Hong Ren,Xiangliang Deng,Xinguo Su
Downregulation of long non-coding RNA MAFG-AS1 represses tumorigenesis of colorectal cancer cells through the microRNA-149-3p-dependent inhibition of HOXB8
Cancer Cell Int. 2020 Oct 19;20:511.;doi: 10.1186/s12935-020-01485-4. (IF 4.175)

87. Qihan Song,Fengli Zhao,Jingfan Yao,Hailin Dai,Lei Hu,Shun Yu
Protective effect of microRNA-134-3p on multiple sclerosis through inhibiting PRSS57 and promotion of CD34 + cell proliferation in rats
J Cell Biochem. 2020 Nov;121(11):4347-4363.;doi: 10.1002/jcb.29643. (IF 4.237)

88. Linbao Wen,Jingwei Sun,Xionggao Chen,Ruili Du
miR-135b-dependent downregulation of S100B promotes neural stem cell differentiation in a hypoxia/ischemia-induced cerebral palsy rat model
AM J PHYSIOL-CELL PH. 2020 Dec 1;319(6):C955-C966.;doi: 10.1152/ajpcell.00481.2019. (IF 3.485)

89. Murong Bao,Gaoxia Liu,Jia Song,Yidan Gao
Long non-coding RNA MALAT1 promotes odontogenic differentiation of human dental pulp stem cells by impairing microRNA-140-5p-dependent downregulation of GIT2
Cell Tissue Res. 2020 Dec;382(3):487-498.;doi: 10.1007/s00441-020-03246-1. (IF 4.044)

90. Xiong Guo,Ling Liu,Qi Zhang,Weiming Yang,Yang Zhang
E2F7 Transcriptionally Inhibits MicroRNA-199b Expression to Promote USP47, Thereby Enhancing Colon Cancer Tumor Stem Cell Activity and Promoting the Occurrence of Colon Cancer
Front Oncol. 2021 Jan 7;10:565449.;doi: 10.3389/fonc.2020.565449. (IF 4.848)

91. Yi Hou,Hai Li,Wei Huo
MicroRNA-495 alleviates ulcerative interstitial cystitis via inactivating the JAK-STAT signaling pathway by inhibiting JAK3
Int Urogynecol J. 2021 Jan 8.;doi: 10.1007/s00192-020-04593-x. (IF 2.071)

92. Xinyu Zeng,Huiqun Liao,Fusen Wang
MicroRNA-384 inhibits nasopharyngeal carcinoma growth and metastasis via binding to Smad5 and suppressing the Wnt/β-catenin axis
Cytotechnology. 2021 Apr;73(2):203-215.;doi: 10.1007/s10616-021-00458-3. (IF 1.777)

93. Meng-Xia Zhu,Lin-Hui Huang,Yi-Ke Zhu,Xing-Jun Cai
LncRNA NEAT1 promotes airway smooth muscle cell inflammation by activating the JAK3/STAT5 pathway through targeting of miR-139
Exp Lung Res. Apr-May 2021;47(4):161-172.;doi: 10.1080/01902148.2021.1876792. (IF 1.702)

94. Zhihua Guo,Huangyang Ye,Xiaobin Zheng,Weiqiang Yin,Jianxing He
Extracellular vesicle-encapsulated microRNA-425-derived from drug-resistant cells promotes non-small-cell lung cancer progression through DAPK1-medicated PI3K/AKT pathway
J Cell Physiol. 2021 May;236(5):3808-3820.;doi: 10.1002/jcp.30126. (IF 5.546)