上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。
产品资料 – Buffers – Buffers
Luciferase Cell Lysis Buffer(已停产且无替代库存售完为止) 收藏
货 号
规 格
价 格(元)
北京库存
上海库存
广州库存
成都库存
苏州库存
#B3321S
25 ml
659.00
无
无
无
无
无
Download:
isoschizomers |
compatible ends |
single letter code
product information
Luciferase Cell Lysis Buffer (LCLB) is a proprietary formulation developed to produce mammalian cell lysates for reporter assays. This lysis buffer is compatible with reagents for assaying the activity of Gaussia as well as other luciferases (e.g. Renilla & Firefly) and β-galactosidase (Figure 1A-D).
Cellartis iPSC Single-Cell Cloning DEF-CS™ Culture Media Kit
1 kit
￥7,860
收藏产品 加入购物车
无标题文档
从开始到结束：人iPS细胞基因编辑和单细胞克隆系统 Start-To-Finish hiPSC Editing and Single-Cell-Cloning Systems
人诱导多能干细胞（Human induced pluripotent stem cells，hiPSCs)是一个功能强大的研究工具和疾病模型，因为其具备增殖、自我更新、分化为多种细胞类型的能力。此外，人iPS细胞还可以再现细胞来源个体的疾病表型和基因型。将人iPS细胞结合基于CRISPR/Cas9的基因编辑技术，可以在同基因的细胞系中研究特定基因改变引起的功能影响，因为基因编辑过的人iPS细胞能提供可再生的、无遗传变异的患病细胞和健康细胞资源。 CRISPR/Cas9系统已经成为一个强大的基因编辑工具，因为其高的靶向特异性，编辑效率和易用性。该技术的强大主要源于它的简单，因为全部只需要一个Cas9核酸酶结合一个单链向导RNA（single guide RNA，sgRNA) 就可以决定靶向特异性（Jinek et al. 2012)。这种RNA编程的（RNA-programmable）方法利用了非同源末端连接(non-homologous end joining，NHEJ) DNA修复途径的易错特性，进而产生基因敲除(通过插入/删除)。这种方法也可以通过同源重组（homology-directed repair，HDR)途径被用于基因敲入。
CRISPR/Cas9系统的组件可以通过多种方法成功导入至靶细胞，包括基于载体的表达系统，RNA转染, 导入Cas9/sgRNA核糖核蛋白（ribonucleoprotein，RNP)复合物(Sander and Joung 2014)等。与基于载体的方法相比，直接导入Cas9/sgRNA RNPs提供了一种快速转入基因编辑实验的方法，同时脱靶效应的可能性更小（Kim et al. 2014)。
一旦Cas9/sgRNA RNP复合物被导入，为了分离和筛选感兴趣的基因型，单细胞必须被分离并扩增为克隆细胞系。传统上,从基因编辑的hiPS细胞建立一个克隆群是低效的、具有挑战性的和耗时的，因为hiPS 细胞是以集落生长和传代的。通常，这会导致细胞死亡和提前分化。然而，Cellartis单细胞克隆系统包含一个成分确定的培养系统(由基础培养基，包被剂，和添加剂组成)，用于有效形成单细胞克隆和扩增基因编辑后的hiPSC克隆。DEF-CS culture system (Asplund et al. 2016)是一个基于单层培养的系统，规避了集落状培养带来的挑战，允许单细胞传代和促进接种的单细胞存活和后续扩增。
Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Code No. Y30021) 是一个完整的，在无饲养层和成分确定条件下，用于hiPSCs有效扩增和扩大生产的系统。试剂盒包含了从接种单细胞至96孔板到扩增至48孔板过程中所有必要的试剂。单细胞克隆扩增后，hiPSCs可以使用Cellartis DEF-CS 500 Culture System (Code No. Y30010)继续培养。
关联基因编辑系统： 一、电穿孔法基因编辑 通过电穿孔的方法对hiPSCs进行有效的基因编辑，Takara提供重组Cas9蛋白质和产生大量sgRNAs所有必要的试剂，用于电穿孔hiPSCs。后续与Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Code No. Y30021)一起使用，用于进行基因编辑后的单细胞克隆形成和扩增。单细胞克隆扩增后，hiPSCs可以使用Cellartis DEF-CS 500 Culture System (Code No. Y30010)继续培养。 二、Gesicle法基因编辑 Gesicles法是一种无毒的、高效的、替代电穿孔的方法，不依赖昂贵的设备或耗材。通过Gesicle法导入Cas9/sgRNA复合物进行有效的基因编辑，Takara提供所有的必要试剂用于产生细胞衍生的纳米囊泡（Gesicles），来转导Cas9和sgRNA至hiPSCs。后续可与Cellartis iPSC Single-Cell Cloning DEF-CS Culture Media Kit (Code No. Y30021)一起使用，用于进行基因编辑后的单细胞克隆形成和扩增。单细胞克隆扩增后，hiPSCs可以使用Cellartis DEF-CS 500 Culture System (Code No. Y30010)继续培养。
■ 产品特点
· Highly efficient gene editing in hiPSCs—use either electroporation or gesicles to deliver Cas9 protein and sgRNA with no genomic integration and reduced off-target effects · Superior single-cell survival—edited hiPSCs exhibit high (typically ~50%) survival when seeded as single cells in a 96-well plate · Maintenance of pluripotency after editing and during single-cell cloning—hiPSCs maintain high levels (>90%) of pluripotency markers Oct-4, TRA-1-60, and SSEA-4 · Stable karyotype from start to finish—maintain a normal and stable karyotype throughout editing, single-cell cloning, and expansion · Flexibility to perform gene editing experiments your way—choose from complete kits that provide editing and single-cell cloning solutions using either electroporation-based or utilize the culture media kit (Code No. Y30021) to perform efficient single-cell cloning following your own editing experiments · Continuous use of DEF-CS technology throughout gene editing/single-cell cloning experiments—use the Cellartis DEF-CS 500 Culture System (Code No. Y30010) before and during gene editing experiments, then during scale-up after single-cell cloning experiments
关联基因编辑产品 电穿孔法基因编辑 Guide-it™ sgRNA In Vitro Transcription Kit (632635; 1 kit) Guide-it™ Recombinant Cas9 (Electroporation-Ready) (632641; 100 μg) Gesicle法基因编辑 Guide-it™ CRISPR/Cas9 Gesicle Production System (632613；1 kit)
■ 产品应用
· CRISPR/Cas9-mediated gene editing of hiPSCs · Single-cell cloning of hiPSCs · Disease modeling
■ 参考文献
Asplund, A. et al. One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells. Stem Cell Rev. Reports12, 90-104 (2016). Jinek, M. et al. A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science337, (2012). Kim, S., Kim, D., Cho, S. W., Kim, J. & Kim, J.-S. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 24, 1012-9 (2014). Sander, J.-D. & Joung, J.-K. CRISPR-Cas9 systems for genomic editing, regulation and targeting. Nat. Biotechnol. 32, 347-55 (2014).
■ 产品详情请点击：
Human induced pluripotent stem cells (hiPSCs) growing in DEF-CS medium were edited with Cas9 and sgRNA specific to CD81. Pluripotency is maintained in edited human induced pluripotent stem cells (hiPSCs) that have been seeded as single cells. 12 clonal lines created from single-seeded hiPSCs exhibit high levels of Oct-4 (97-99%), TRA-1-60 (98-99%), and SSEA-4 (98-99%).
1.Potential circadian effects on translational failure for neuroprotection，Nature,2020,582, 395–398
Epigenetic therapy inhibits metastases by disrupting premetastatic niches.Nature,2020,doi.org/10.1038/s41586-020-2054-x
2.Chimeric peptidomimetic antibiotics against Gram-negative bacteria，Nature,2019,doi:10.1038/s41586-019-1665-6
PU.1 controls fibroblast polarization and tissue fibrosis.Nature,2019,566,344–349
3.Mitochondrial unfolded protein response controls matrix pre-RNA processing and translation,Nature,2016,534,710-713
4.Transfer of mitochondria from astrocytes to neurons after stroke.Nature,2016,535,551-555
5.Signalling thresholds and negative B-cell selection in acute lymphoblastic leukaemia.Nature,2015, 521,357–361
6.The sonic hedgehog factor GLI1 imparts drug resistance through inducible glucuronidation.Nature,2014,511(7507),90-93
7.The sonic hedgehog factor GLI1 imparts drug resistance through inducible glucuronidation.Nature,2014,511(7507),90-93
8.Sema3A regulates bone-mass accrual through sensory innervations,Nature,2013,497(7450),490-493
9.Acetylation-dependent regulation of endothelial Notch signalling by the SIRT1 deacetylase.Nature,2011,473(7346),234-238
10.Embryonic lethal phenotype reveals a function of TDG in maintaining epigenetic stability.Nature,2011,470(7334),419-423
11.Intravenous delivery of a multi-mechanistic cancer-targeted oncolytic poxvirus in humans.Nature,2011,477(7362),99-102
12.Frequent inactivation of A20 in B-cell lymphomas.Nature,2009,459(7247),712-716
13.Modulation of microRNA processing by p53.Nature,2009,460(7254),529-33
14.Oncogenic mutations of ALK kinase in neuroblastoma.Nature,2008,455(7215),971-4
15.Hyaluronic acid–bilirubin nanomedicine for targeted modulation of dysregulated intestinal barrier, microbiome and immune responses in colitis.Nature Materials,2019,doi: 10.1038/s41563-019-0462-9
16.Parenchymal and stromal tissue regeneration of tooth organ by pivotal signals reinstated in decellula….Nature Materials,2019,18(6),627-637
17.Guiding intracortical brain tumour cells to an extracortical cytotoxic hydrogel using aligned polymer….Nature Materials,2014,13(3),308-16
18.A Bacterial Effector Reveals the V-ATPase-ATG16L1 Axis that Initiates Xenophagy.Cell,2019,178(3),552-566
19.Higher-Order Clustering of the Transmembrane Anchor of DR5 Drives Signaling.Cell,2019,176(6),1477-1489
20.Insulin Receptor Associates with Promoters Genome-wide and Regulates Gene Expression.Cell,2019,177(3),722-736
21.B-Cell-Specific Diversion of Glucose Carbon Utilization Reveals a Unique Vulnerability in B Cell Mali….Cell,2018,173(2),470-484
22.Fusobacterium nucleatum Promotes Chemoresistance to Colorectal Cancer by Modulating Autophagy
Cell,2017,170(3),548-563
23.Methyltransferase SETD2-Mediated Methylation of STAT1 Is Critical for Interferon Antiviral Activity
Cell,2017,170(3),492-506
24.Loss of 5-Hydroxymethylcytosine Is an Epigenetic Hallmark of Melanoma.Cell,2012,150(6),1135-1146
25.Mapping the Hallmarks of Lung Adenocarcinoma with Massively Parallel Sequencing.Cell,2012,150(6),1107-1120
26.Iron-Export Ferroxidase Activity of β-Amyloid Precursor Protein Is Inhibited by Zinc in Alzheimers D….Cell,2010,142(6),857-867
27.Autocrine VEGF Signaling Is Required for Vascular Homeostasis.Cell,2007,130(4),691-703
28.Genetic risk of extranodal natural killer T-cell lymphoma: a genome-wide association study in multipl…
Lancet Oncology,2019,pii: S1470-2045(19)30799-5
29.Precise targeting of?POLR2A?as a therapeutic strategy for human triple negative breast cancer.Nature Nanotechnology,2019,14 , 388–397
30.Metal nanoparticles in the presence of lipopolysaccharides trigger the onset of metal allergy in mice…
Nature Nanotechnology,2016,11,808–816
31.Chemical modification of PS-ASO therapeutics reduces cellular protein-binding and improves the therap…
Nature Biotechnology,2019,37640–650
32.Multi-omic measurements of heterogeneity in HeLa cells across laboratories
Nature Biotechnology,2019,37,314–322
33.Dual effects of carbon monoxide on pericytes and neurogenesis in traumatic brain injury.
