DDR1 Knockout HEK293T RIPA Lysate(DDR1基因敲除HEK293T细胞RIPA裂解液)是通过同时表达Cas9、目的基因sgRNA和puromycin抗性基因并实现了目的基因CRISPR敲除的多克隆HEK293T细胞的RIPA裂解液。该细胞中目的基因的敲除已经通过T7EI法的验证。本产品可用于该目的基因敲除后其信号通路相关蛋白的研究,也可以用于该基因相应抗体的验证。
本RIPA裂解液源于可同时表达Cas9、目的基因sgRNA和puromycin抗性基因的慢病毒感染HEK293T细胞并经过puromycin筛选后获得的多克隆细胞。制备该细胞的相应慢病毒的基因序列的关键图谱信息请参考图1。
图1. 可同时表达sgRNA、Cas9和puromycin抗性的慢病毒其基因序列的关键图谱信息。
该细胞中目的基因的敲除已经通过T7EI法的验证。
由于本细胞是通过CRISPR/Cas9技术获得的多克隆细胞,基于CRISPR/Cas9技术的特点,理论上平均有2/3的细胞发生移码突变而导致了目的基因的敲除,平均有1/3的细胞并未发生移码突变。很多情况下有约2/3的细胞发生目的基因的敲除,已经足以进行很多的目的基因的生物学功能的研究了。如果希望获得100%基因敲除的细胞,可以自行使用本产品筛选单克隆细胞,或者委托碧云天进行单克隆细胞株的筛选服务。
本RIPA裂解液用于实验时,建议同时选购无任何靶向的对照细胞RIPA裂解液Control Knockout HEK293T RIPA Lysate (L00025)或靶向GFP的对照RIPA裂解液GFP Knockout HEK293T RIPA Lysate (L00027)。
碧云天同时提供基于CRISPR/Cas9技术的DDR1基因敲除的质粒(L15445 pLenti-DDR1-sgRNA)、慢病毒(L15446 DDR1 Knockout Lentivirus)、HEK293T细胞(L15447 DDR1 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L15448 DDR1 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L15449 DDR1 Knockout HEK293T Trizol Lysate)等产品,具体请在碧云天网站查询或在本产品网页点击相应产品。
用于制备本产品的RIPA裂解液为碧云天生产的P0013B RIPA裂解液(强),其具体组分请查阅产品网页或说明书。
DDR1基因的基本信息如下:
Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
Human | DDR1 | 780 | BC070070 | NM_013994 |
About the gene | |
Official Symbol | DDR1 |
Previous Symbol | NTRK4|PTK3A|NEP|CAK|EDDR1 |
Official Full Name | discoidin domain receptor tyrosine kinase 1 |
Synonyms | RTK6; CD167 |
Location | 6p21.33 |
Gene Type | protein-coding gene |
Uniprot ID | Q08345 |
Pathway/Library | Kinases Library |
Gene Summary | Receptor tyrosine kinases (RTKs) play a key role in the communication of cells with their microenvironment. These molecules are involved in the regulation of cell growth, differentiation and metabolism. The protein encoded by this gene is a RTK that is widely expressed in normal and transformed epithelial cells and is activated by various types of collagen. This protein belongs to a subfamily of tyrosine kinase receptors with a homology region to the Dictyostelium discoideum protein discoidin I in their extracellular domain. Its autophosphorylation is achieved by all collagens so far tested (type I to type VI). In situ studies and Northern-blot analysis showed that expression of this encoded protein is restricted to epithelial cells, particularly in the kidney, lung, gastrointestinal tract, and brain. In addition, this protein is significantly over-expressed in several human tumors from breast, ovarian, esophageal, and pediatric brain. This gene is located on chromosome 6p21.3 in proximity to several HLA class I genes. Alternative splicing of this gene results in multiple transcript variants. |
产品编号 | 产品名称 | 包装 |
L15448 | DDR1 Knockout HEK293T RIPA Lysate | 100µg |
— | 说明书 | 1份 |
-20℃保存,3个月有效;-80℃保存,至少一年有效。
注意事项:碧云天拥有sgRNA序列的知识产权,如果需要sgRNA序列,请在订购后发送邮件向info@beyotime.com索取。sgRNA序列信息与本产品,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。使用者在发表研究论文或结果时,应注明来源。
对于非目录产品的CRISPR基因敲除细胞RIPA裂解液的定制,可联系碧云天技术服务service@beyotime.com。
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。
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产品编号 | 产品名称 | 包装 |
L00025 | Control Knockout HEK293T RIPA Lysate | 100µg |
L00027 | GFP Knockout HEK293T RIPA Lysate | 100µg |
P0009 | BCA蛋白浓度测定试剂盒(增强型) | 5000次 |
P0010 | BCA蛋白浓度测定试剂盒(增强型) | 500次 |
P0010S | BCA蛋白浓度测定试剂盒(增强型) | 200次 |
P0011 | BCA蛋白浓度测定试剂盒 | 5000次 |
P0012 | BCA蛋白浓度测定试剂盒 | 500次 |
P0012S | BCA蛋白浓度测定试剂盒 | 200次 |
P0012A | SDS-PAGE凝胶配制试剂盒 | 可制30-50块胶 |
P0012AC | SDS-PAGE凝胶快速配制试剂盒 | 可制30-50块胶 |
P0013B | RIPA裂解液(强) | 100ml |
P0014A/B | SDS-PAGE电泳液(Tris-Gly, Powder) | 1/10L |
P0014C/D | SDS-PAGE电泳液(Tris-Gly, 10X) | 100/500ml |
P0015 | SDS-PAGE蛋白上样缓冲液(5X) | 2ml |
P0015L | SDS-PAGE蛋白上样缓冲液(5X) | 15ml |
P0015B | SDS-PAGE蛋白上样缓冲液(2X) | 5ml |
P0015F | SDS-PAGE蛋白上样缓冲液(6X) | 2ml |