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TRIM67基因敲除HEK293T细胞RIPA裂解液
产品编号: L10978
产品包装:100µg
价格: ¥ 1258.00
产品简介
使用说明
产品文件
相关产品
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产品编号 产品名称 产品包装 产品价格
L10975 TRIM67基因敲除质粒 5µg 999.00元
L10976 TRIM67基因敲除慢病毒 10^8 TU 6999.00元
L10977 TRIM67基因敲除HEK293T细胞 1支/瓶 7999.00元
L10978 TRIM67基因敲除HEK293T细胞RIPA裂解液 100µg 1258.00元
L10979 TRIM67基因敲除HEK293T细胞Trizol裂解液 500µl 1258.00元

TRIM67 Knockout HEK293T RIPA Lysate(TRIM67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TRIM67基因敲除的质粒(L10975 pLenti-TRIM67-sgRNA)慢病毒(L10976 TRIM67 Knockout Lentivirus)HEK293T细胞(L10977 TRIM67 Knockout HEK293T Cells)HEK293T敲除细胞的RIPA裂解液(L10978 TRIM67 Knockout HEK293T RIPA Lysate)HEK293T敲除细胞的Trizol裂解液(L10979 TRIM67 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。

TRIM67基因的基本信息如下:

Species Gene Symbol Gene ID GenBank Accession Transcript
Human TRIM67 440730 - NM_001004342

 

About the gene
Official Symbol TRIM67
Previous Symbol -
Official Full Name tripartite motif containing 67
Synonyms TNL
Location 1q42.2
Gene Type protein_coding
Uniprot ID Q6ZTA4.3
Pathway/Library Ubiquitin Ligases Genes Library
Gene Summary Tripartite motif (TRIM) proteins have been increasingly appreciated as important antiviral factors that suppress the replication of a wide range of RNA and DNA viruses. TRIM proteins inhibit viral replication by either directly targeting viral components or modulating innate immune responses that result in antiviral gene expression. For example, TRIM19 (also known as promyelocytic leukemia protein (PML)) restricts multiple RNA viruses and DNA viruses, including herpesviruses, by organizing PML nuclear bodies. Several TRIM proteins that suppress KSHV reactivation. TRIM67, was commonly silenced in colorectal cancer and its downregulation was associated with poor survival. Trim67 knockout in ApcMin/+ mice increased the incidence, multiplicity, and burden of colorectal tumors. Similarly, colon-specific knockout of Trim67 significantly accelerated azoxymethane-induced colorectal cancer in mice. RNA sequencing revealed that the antitumor effect of TRIM67 was mediated by activation of the p53 signaling pathway. TRIM67 interacted directly with the C-terminus of p53, inhibiting p53 degradation by its ubiquitin ligase MDM2. TRIM67 was also a transcriptional target of p53; upon cellular stress, p53 bound to the TRIM67 promoter and induced significant upregulation of TRIM67, thereby forming a TRIM67/p53 self-amplifying loop that boosts p53-induced cell growth inhibition and apoptosis. Consequently, loss of this p53-positive regulatory program profoundly compromised p53-mediated responses to chemotherapy-induced DNA damage. Dampened p53 response was also observed in tumors of Trim67 knockout mice and Trim67 knockout embryonic fibroblasts. TRIM67 reactivation restored p53 activation and sensitized colorectal cancer cells to chemotherapy in vitro and in vivo. TRIM67 thus functions as a pivotal tumor suppressor in colorectal cancer and is a potential target for improving chemotherapy responsiveness.
包装清单:
产品编号 产品名称 包装
L10978 TRIM67 Knockout HEK293T RIPA Lysate 100µg
说明书 1份
保存条件:

-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。

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. 细胞RIPA裂解液样品的蛋白浓度测定:
推荐使用碧云天生产的BCA蛋白浓度测定试剂盒(P0009/P0010/P0010S/P0011/P0012/P0012S)测定蛋白浓度。由于本产品含有较高浓度的去垢剂,不建议使用Bradford法测定本产品的蛋白浓度。RIPA裂解液的使用说明请参考:https://www.beyotime.com/product/P0013B.htm
2. 细胞RIPA裂解液的使用:
本RIPA裂解液测定浓度后加入SDS-PAGE蛋白上样缓冲液即可用于SDS-PAGE、Western检测、以及目的基因抗体的验证等。
相关产品:
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
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