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| 产品目录号 | GKC-LMP010097 | ||
| 产品名称 | Human MAP2K6-A549 KO Cell Pool | ||
| 基因编号 | MAP2K6 | ||
| Uniprot_id | P52564 | ||
| 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 | Dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. With MAP3K3/MKK3, catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in the MAP kinases p38 MAPK11, MAPK12, MAPK13 and MAPK14 and plays an important role in the regulation of cellular responses to cytokines and all kinds of stresses. Especially, MAP2K3/MKK3 and MAP2K6/MKK6 are both essential for the activation of MAPK11 and MAPK13 induced by environmental stress, whereas MAP2K6/MKK6 is the major MAPK11 activator in response to TNF. MAP2K6/MKK6 also phosphorylates and activates PAK6. The p38 MAP kinase signal transduction pathway leads to direct activation of transcription factors. Nuclear targets of p38 MAP kinase include the transcription factors ATF2 and ELK1. Within the p38 MAPK signal transduction pathway, MAP3K6/MKK6 mediates phosphorylation of STAT4 through MAPK14 activation, and is therefore required for STAT4 activation and STAT4-regulated gene expression in response to IL-12 stimulation. The pathway is also crucial for IL-6-induced SOCS3 expression and down-regulation of IL-6-mediated gene induction; and for IFNG-dependent gene transcription. Has a role in osteoclast differentiation through NF-kappa-B transactivation by TNFSF11, and in endochondral ossification and since SOX9 is another likely downstream target of the p38 MAPK pathway. MAP2K6/MKK6 mediates apoptotic cell death in thymocytes. Acts also as a regulator for melanocytes dendricity, through the modulation of Rho family GTPases. | ||
| 细胞系生成 | 采用CRISPR方法生成Human MAP2K6-A549 KO Cell Pool | ||
| 数据说明 | Sanger 测序结果显示Human MAP2K6-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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