NLK Knockout HEK293T RIPA Lysate(NLK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NLK基因敲除的质粒(L16180 pLenti-NLK-sgRNA)、慢病毒(L16181 NLK Knockout Lentivirus)、HEK293T细胞(L16182 NLK Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L16183 NLK Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L16184 NLK 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。
NLK基因的基本信息如下：
| Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
| Human | NLK | 51701 | BC064663 | NM_016231 |
| About the gene | |
| Official Symbol | NLK |
| Previous Symbol | - |
| Official Full Name | nemo like kinase |
| Synonyms | - |
| Location | 17q11.2 |
| Gene Type | protein-coding gene |
| Uniprot ID | Q9UBE8 |
| Pathway/Library | Kinases Library |
| Gene Summary | Serine/threonine-protein kinase that regulates a number of transcription factors with key roles in cell fate determination. Positive effector of the non-canonical Wnt signaling pathway, acting downstream of WNT5A, MAP3K7/TAK1 and HIPK2. Activation of this pathway causes binding to and phosphorylation of the histone methyltransferase SETDB1. The NLK-SETDB1 complex subsequently interacts with PPARG, leading to methylation of PPARG target promoters at histone H3K9 and transcriptional silencing. The resulting loss of PPARG target gene transcription inhibits adipogenesis and promotes osteoblastogenesis in mesenchymal stem cells (MSCs). Negative regulator of the canonical Wnt/beta-catenin signaling pathway. Binds to and phosphorylates TCF7L2/TCF4 and LEF1, promoting the dissociation of the TCF7L2/LEF1/beta-catenin complex from DNA, as well as the ubiquitination and subsequent proteolysis of LEF1. Together these effects inhibit the transcriptional activation of canonical Wnt/beta-catenin target genes. Negative regulator of the Notch signaling pathway. Binds to and phosphorylates NOTCH1, thereby preventing the formation of a transcriptionally active ternary complex of NOTCH1, RBPJ/RBPSUH and MAML1. Negative regulator of the MYB family of transcription factors. Phosphorylation of MYB leads to its subsequent proteolysis while phosphorylation of MYBL1 and MYBL2 inhibits their interaction with the coactivator CREBBP. Other transcription factors may also be inhibited by direct phosphorylation of CREBBP itself. Acts downstream of IL6 and MAP3K7/TAK1 to phosphorylate STAT3, which is in turn required for activation of NLK by MAP3K7/TAK1. Upon IL1B stimulus, cooperates with ATF5 to activate the transactivation activity of C/EBP subfamily members. Phosphorylates ATF5 but also stabilizes ATF5 protein levels in a kinase-independent manner (PubMed:25512613). NLK_HUMAN,Q9UBE8 |
| 产品编号 | 产品名称 | 包装 |
| L16183 | NLK 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。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
| 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 |
