SRPK2 Knockout HEK293T RIPA Lysate(SRPK2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SRPK2基因敲除的质粒(L16565 pLenti-SRPK2-sgRNA)、慢病毒(L16566 SRPK2 Knockout Lentivirus)、HEK293T细胞(L16567 SRPK2 Knockout HEK293T Cells)、HEK293T敲除细胞的RIPA裂解液(L16568 SRPK2 Knockout HEK293T RIPA Lysate)、HEK293T敲除细胞的Trizol裂解液(L16569 SRPK2 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。
SRPK2基因的基本信息如下:
Species | Gene Symbol | Gene ID | GenBank Accession | Transcript |
Human | SRPK2 | 6733 | BC035214 | NM_182691 |
About the gene | |
Official Symbol | SRPK2 |
Previous Symbol | - |
Official Full Name | SRSF protein kinase 2 |
Synonyms | SFRSK2 |
Location | 7q22.3 |
Gene Type | protein-coding gene |
Uniprot ID | P78362 |
Pathway/Library | Kinases Library |
Gene Summary | Serine/arginine-rich protein-specific kinase which specifically phosphorylates its substrates at serine residues located in regions rich in arginine/serine dipeptides, known as RS domains and is involved in the phosphorylation of SR splicing factors and the regulation of splicing (PubMed:9472028, PubMed:18559500, PubMed:21056976). Promotes neuronal apoptosis by up-regulating cyclin-D1 (CCND1) expression (PubMed:19592491). This is done by the phosphorylation of SRSF2, leading to the suppression of p53/TP53 phosphorylation thereby relieving the repressive effect of p53/TP53 on cyclin-D1 (CCND1) expression (PubMed:21205200). Phosphorylates ACIN1, and redistributes it from the nuclear speckles to the nucleoplasm, resulting in cyclin A1 but not cyclin A2 up-regulation (PubMed:18559500). Plays an essential role in spliceosomal B complex formation via the phosphorylation of DDX23/PRP28 (PubMed:18425142). Probably by phosphorylating DDX23, leads to the suppression of incorrect R-loops formed during transcription; R-loops are composed of a DNA:RNA hybrid and the associated non-template single-stranded DNA (PubMed:28076779). Can mediate hepatitis B virus (HBV) core protein phosphorylation (PubMed:12134018). Plays a negative role in the regulation of HBV replication through a mechanism not involving the phosphorylation of the core protein but by reducing the packaging efficiency of the pregenomic RNA (pgRNA) without affecting the formation of the viral core particles (PubMed:16122776). SRPK2_HUMAN,P78362 |
产品编号 | 产品名称 | 包装 |
L16568 | SRPK2 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 |