数据库共享平台 -- 细胞查询-- 详细信息
       
产品目录号  GKC-LMP010043
产品名称  Human CSNK2A1-A549 KO Cell Pool
基因编号   CSNK2A1
Uniprot_id  P68400
宿主细胞   A549 
组织来源   人非小细胞肺癌细胞
规格   1×106cells/T25培养瓶或1×106cells/冻存管
培养基  MEM+10%FBS+1%P/S
筛选标记   N/A
生长特性   贴壁细胞,上皮细胞样
培养条件   37℃,5% CO2的培养箱
传代比例   1/2到1/3传代,2-3天长满
筛选标记   N/A
换液频率   2-3天换液
支原体检测结果   阴性
蛋白组验证结果   N/A
抗体验证结果   N/A
目标基因介绍   Catalytic subunit of a constitutively active serine/threonine-protein kinase complex that phosphorylates a large number of substrates containing acidic residues C-terminal to the phosphorylated serine or threonine (PubMed:11239457, PubMed:11704824, PubMed:16193064, PubMed:19188443, PubMed:20625391, PubMed:22406621, PubMed:24962073, PubMed:31439799). Regulates numerous cellular processes, such as cell cycle progression, apoptosis and transcription, as well as viral infection (PubMed:12631575, PubMed:19387552, PubMed:19387551). May act as a regulatory node which integrates and coordinates numerous signals leading to an appropriate cellular response (PubMed:12631575, PubMed:19387552, PubMed:19387551). During mitosis, functions as a component of the p53/TP53-dependent spindle assembly checkpoint (SAC) that maintains cyclin-B-CDK1 activity and G2 arrest in response to spindle damage (PubMed:11704824, PubMed:19188443). Also required for p53/TP53-mediated apoptosis, phosphorylating 'Ser-392' of p53/TP53 following UV irradiation. Can also negatively regulate apoptosis (PubMed:11239457). Phosphorylates the caspases CASP9 and CASP2 and the apoptotic regulator NOL3 (PubMed:16193064). Phosphorylation protects CASP9 from cleavage and activation by CASP8, and inhibits the dimerization of CASP2 and activation of CASP8 (PubMed:16193064). Phosphorylates YY1, protecting YY1 from cleavage by CASP7 during apoptosis (PubMed:22184066). Regulates transcription by direct phosphorylation of RNA polymerases I, II, III and IV (PubMed:19387550, PubMed:12631575, PubMed:19387552, PubMed:19387551, PubMed:23123191). Also phosphorylates and regulates numerous transcription factors including NF-kappa-B, STAT1, CREB1, IRF1, IRF2, ATF1, ATF4, SRF, MAX, JUN, FOS, MYC and MYB (PubMed:19387550, PubMed:12631575, PubMed:19387552, PubMed:19387551, PubMed:23123191). Phosphorylates Hsp90 and its co-chaperones FKBP4 and CDC37, which is essential for chaperone function (PubMed:19387550). Mediates sequential phosphorylation of FNIP1, promoting its gradual interaction with Hsp90, leading to activate both kinase and non-kinase client proteins of Hsp90 (PubMed:30699359). Regulates Wnt signaling by phosphorylating CTNNB1 and the transcription factor LEF1 (PubMed:19387549). Acts as an ectokinase that phosphorylates several extracellular proteins (PubMed:19387550, PubMed:12631575, PubMed:19387552, PubMed:19387551). During viral infection, phosphorylates various proteins involved in the viral life cycles of EBV, HSV, HBV, HCV, HIV, CMV and HPV (PubMed:19387550, PubMed:12631575, PubMed:19387552, PubMed:19387551). Phosphorylates PML at 'Ser-565' and primes it for ubiquitin-mediated degradation (PubMed:20625391, PubMed:22406621). Plays an important role in the circadian clock function by phosphorylating BMAL1 at 'Ser-90' which is pivotal for its interaction with CLOCK and which controls CLOCK nuclear entry (By similarity). Phosphorylates CCAR2 at 'Thr-454' in gastric carcinoma tissue (PubMed:24962073). Phosphorylates FMR1, promoting FMR1-dependent formation of a membraneless compartment (PubMed:30765518, PubMed:31439799).
细胞系生成   采用CRISPR方法生成Human CSNK2A1-A549 KO Cell Pool
数据说明   Sanger 测序结果显示Human CSNK2A1-A549 KO Cell Pool敲除效率为100%
应用   体内和体外测定
复苏   1)在37℃水浴中预热完全培养基。 2)将冻存管在 37℃水浴中解冻1-2分钟。 3)将冻存管转移到生物安全柜中,并用70%乙醇擦拭表面。 4)拧开冻存管管盖,将细胞悬液轻轻转移到含有9mL完全培养基的无菌离心管中。 5)在室温下以125g离心5-7分钟,弃上清。 6)用5mL的完整培养基重悬细胞沉淀,将细胞悬液转移到T25培养瓶中。 7)将细胞转移到37℃,5% CO2的培养箱中培养。 8)参考传代比例:1/2到1/3传代,2-5天长满。
传代   1)待培养瓶中细胞汇合度至80%-90%以上,可进行细胞传代。 2)将培养基、PBS、胰酶(0.25%Trypsin_EDTA Gibco 25200-056)等从4℃冰箱中拿出,置于37℃水浴中温度接近37℃时取出并在瓶子表面喷洒75%酒精后置于生物安全柜中。 3)从培养箱中取出待传代的培养瓶,瓶身喷洒75%酒精后置于生物安全柜中。 4)为避免冲散细胞,沿培养瓶上壁PBS润洗细胞,清洗细胞后弃去,T25加2mL。 5)加入对应体积的胰酶(T75加1.5mL,T25加0.5mL),并轻轻晃动瓶身使胰酶平铺满细胞底部。可根据实际情况适当增加或减少用量。约1-2min后大部分细胞脱落时,加入对应体积的完全培养基终止消化,并用5mL移液管轻轻吹打至细胞全部脱落。 6)将细胞悬液转移至15mL离心管,悬液300g离心5min,弃上清。 7)移取5mL完全培养基重悬细胞,按需求调整接种比例,并补充培养瓶中完全培养基,T75加至13-15mL,T25加至5mL,加1%双抗。 8)盖上瓶盖拧紧后轻轻晃动瓶身,使细胞混合均匀后置于37℃,5% CO4培养箱中。
细胞冻存   1)准备冻存液,并提前预冷。 2)确保待冻存的细胞满足冻存要求,用显微镜检查以下状态:健康的外观及形态特征、所处生长周期(对数晚期)、无污染或衰退迹象。 3)对细胞进行消化及离心处理(具体步骤参考传代培养流程) 4)按照每管1mL的量添加冻存液重悬细胞,吹打均匀后分装至冻存管。 5)将细胞放在程序降温盒中,在-80℃冰箱中冷冻。 6)后续将细胞转移到液氮罐中,以便长期储存。
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