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报告基因质粒

pNFκB-luc (报告基因质粒)
产品编号: D2206-1μg
产品包装: 1μg
产品价格: 888.00元
产品简介
使用说明
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产品编号 产品名称 产品包装 产品价格
D2206-1μg pNFκB-luc (报告基因质粒) 1μg 888.00元

      pNFκB-luc (报告基因质粒)是碧云天自行研发的用于检测NFκB转录活性水平的报告基因质粒。pNFκB-luc是以碧云天的pGL6质粒为模板,在其多克隆位点插入了多个NFκB结合位点,可以高灵敏度地检测NFκB的激活水平。
      pGL6质粒是用于在哺乳动物细胞中进行萤火虫萤光素酶(firefly luciferase)报告基因检测的新一代质粒。该报告基因质粒比Promega公司的pGL3系列有了全面的改进,一方面对于luciferase的编码进行了改进,确保能更好地在哺乳动物细胞中进行表达,同时对整个质粒中所有可以被预测出的可能的转录因子结合位点全部进行了适当的突变处理,在保持原有功能不变的情况下,使各种转录因子在质粒上的非特异性结合降到最低。
      萤光素、萤光素酶、萤火虫萤光素酶和海肾萤光素酶也经常被称作荧光素、荧光素酶、萤火虫荧光素酶和海肾荧光素酶。
      pNFκB-luc质粒的主要信息如下:

Base pairs 5610
NFκB response element (κB) 26-65
luc2 reporter gene 111-1763
SV40 late poly(A) signal 1798-2019
SV40 early enhancer/promoter 2067-2485
Synthetic neomycin phosphotransferase
(Neor) coding region 2510-3304
Synthetic poly(A) signal 3329-3377
Reporter Vector primer 4 (RVprimer4) binding region 3444-3463
ColE1-derived plasmid replication origin 3701
Synthetic Beta-lactamase (Ampr) coding region 4492-5352
Synthetic poly(A) signal/transcriptional pause site 5457-5610
Reporter Vector primer 3 (RVprimer3) binding region 5559-5578

      pNFκB-luc质粒的图谱如下:

      pNFκB-luc的多克隆位点及NFκB response element的详细图谱如下:

BglI KpnI NheI NFkB response elememt
1 GGCCTAACTG GCCGGTACCG
CTAGC GGGAA
TTTCCGGGAA
TTTCCGGGAA
CCGGATTGAC CGGCCATGGC
GATCG CCCTT
AAAGGCCCTT
AAAGGCCCTT
BglII HindIII
51 TTTCCGGGAA
TTTCC AGATC
TGCAGAAGCT TGGCAATCCG GTACTGTTGG
AAAGGCCCTT
AAAGG TCTAG
ACGTCTTCGA ACCGTTAGGC CATGACAACC

      pNFκB-luc中没有的酶切位点(Restriction enzymes that do not cut pNFκB-luc))包括:

Aat II Afl II Asc I Ase I Bsa I BsaA I BsiW I BspM II
BssH II Eco72 I EcoR I EcoR V Mlu I Nde I Nru I PaeR7 I
PflM I Pme I Pml I Psp1406 I PspA I Rsr II Sac I Sma I
SnaB I Spl I Srf I Tth111 I Vsp I Xcm I Xho I Xma I

      pNFκB-luc中的单酶切位点(Restriction enzymes that cut pNFκB-luc once)包括:

Sfi I GGCCN,NNN`NGGCC 9 BstB I TT`CG,AA 3380
Bgl I GCCN,NNN`NGGC 9 Sal I G`TCGA,C 3394
Acc65 I G`GTAC,C 15 Afl III A`CRYG,T 3644
Asp718 G`GTAC,C 15 ApaL I G`TGCA,C 3958
Kpn I G,GTAC`C 19 HgiE II ACCNNNNNNGGT -1/13 4223
Nhe I G`CTAG,C 21 Not I GC`GGCC,GC 4464
Bgl II A`GATC,T 67 BstX I CCAN,NNNN`NTGG 4488
Hind III A`AGCT,T 77 BstE II G`GTNAC,C 4491
BsrG I T`GTAC,A 602 Ahd I GACNN,N`NNGTC 4566
Dra III CAC,NNN`GTG 1258 Bsu36 I CC`TNA,GG 4922
Gsu I CTGGAG 21/19 1491 Pvu I CG,AT`CG 4936
Bpm I CTGGAG 22/20 1492 Sac II CC,GC`GG 4960
Mun I C`AATT,G 1938 Bst1107 I GTA|TAC 5076
BamH I G`GATC,C 2031 Xca I GTA|TAC 5076
Stu I AGG|CCT 2464 Spe I A`CTAG,T 5395
EcoN I CCTNN`N,NNAGG 2985 BsmA I GTCTC`/9 5407
BsiC I TT`CG,AA 3380 BsmB I CGTCTC 7/11 5408

      pNFκB-luc质粒中推荐使用的测序引物序列如下:
      RVprimer3(5559-5578):
      CTA GCA AAA TAG GCT GTC CC
      pNFκB-luc的全序列信息:D2206-pNFkappaB-luc-seq.txt
包装清单:

产品编号 产品名称 包装
D2206-1μg pNFκB-luc (报告基因质粒) 1μg
D2206-100μg pNFκB-luc (报告基因质粒) 100μg
说明书 1份

保存条件:
      -20℃保存。
注意事项:
      本质粒未经碧云天书面许可不得用于任何商业用途,也不得移交给订货人所在实验室外的任何个人或单位。
      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. 首次使用1μg包装的本产品时,请先取少量本质粒转化大肠杆菌,进行质粒小量、中量或大量抽提后再用于后续用途。抽提获得的质粒可以通过酶切电泳进行鉴定,或通过测序进行鉴定。
2. 100μg包装的本产品质粒浓度为0.1μg/μl,共1ml。可以直接用于酶切或者转染细胞。
3. pNFκB-luc可以用常规的细胞转染方法转染细胞。检测时可以采用碧云天的萤火虫萤光素酶报告基因检测试剂盒(RG005/RG006)或双萤光素酶报告基因检测试剂盒(RG027/RG028)。
4. TNF-α、IL-1β和LPS等是常见的可以激活NFκB的试剂,可以用作pNFκB-luc报告基因检测时的阳性对照。

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