产品分类
ZDHHC3 Knockout K562 Cell Pool
- 产品描述
- 细胞复苏
- 细胞传代
- 细胞冻存
- 抗体验证结果
-
- 品牌: ELEMok138cn太阳集团529
- 商品名称: ZDHHC3 Knockout K562 Cell Pool
- 商品编号: LM01012163812
- Gene Symbol: ZDHHC3
- Ensembl ID: ENSG00000163812
- Uniprot ID: Q9NYG2-2
- 宿主细胞 / 类型: K562/人慢性髓原白血病细胞
- NCBI Gene ID: 51304
- 规格: 1×10^6 cells/ 冻存管
- 筛选标记: N/A
- 生长特性: 悬浮细胞
- 培养条件: 37℃,5% CO2 的培养箱，1/2 到 1/4 传代
- 倍增时间: ~24-36 hours
- 生长培养基: RPMI-1640+10% FBS+1% P/S
- 参考换液频率: 2~3次/周
- 支原体检测结果: 阴性
- 敲除效率（Sanger测序）: >70%
- 蛋白质组验证结果: N/A
- 抗体货号: 添加中
- 目标基因介绍: Golgi-localized palmitoyltransferase that catalyzes the addition of palmitate onto various protein substrates (PubMed:19001095, PubMed:21926431, PubMed:22240897, PubMed:23034182, PubMed:22314500). Has no stringent fatty acid selectivity and in addition to palmitate can also transfer onto target proteins myristate from tetradecanoyl-CoA and stearate from octadecanoyl-CoA (By similarity). Plays an important role in G protein-coupled receptor signaling pathways involving GNAQ and potentially other heterotrimeric G proteins by regulating their dynamic association with the plasma membrane (PubMed:19001095). Palmitoylates ITGA6 and ITGB4, thereby regulating the alpha-6/beta-4 integrin localization, expression and function in cell adhesion to laminin (PubMed:22314500). Plays a role in the TRAIL-activated apoptotic signaling pathway most probably through the palmitoylation and localization to the plasma membrane of TNFRSF10A (PubMed:22240897). In the brain, by palmitoylating the gamma subunit GABRG2 of GABA(A) receptors and regulating their postsynaptic accumulation, plays a role in synaptic GABAergic inhibitory function and GABAergic innervation. Palmitoylates the neuronal protein GAP43 which is also involved in the formation of GABAergic synapses. Palmitoylates NCDN thereby regulating its association with endosome membranes. Probably palmitoylates PRCD and is involved in its proper localization within the photoreceptor. Could mediate the palmitoylation of NCAM1 and regulate neurite outgrowth. Could palmitoylate DNAJC5 and regulate its localization to Golgi membranes. Also constitutively palmitoylates DLG4. May also palmitoylate SNAP25. Could palmitoylate the glutamate receptors GRIA1 and GRIA2 but this has not been confirmed in vivo (By similarity). Could also palmitoylate the D(2) dopamine receptor DRD2 (PubMed:26535572).||May also function as a calcium transporter.
- 细胞开发路径: 采用CRISPR-RNP方法生成稳定KO Cell Pool；Sanger 测序结果显示KO Cell Pool敲除效率>70%。
- 应用: 高敲除效率的基因敲除细胞池（KO Cell Pool），特别适用于初步功能分析、复杂疾病模型的开发、精准药物筛选以及广泛的基因发现研究。KO pool能够无需繁琐的单克隆挑选过程，直接应用于多种类型的测定和分析，大幅提升实验效率。
-
01. 在 37℃水浴中预热完全培养基。
02. 将冻存管在 37℃水浴中解冻 1-2 分钟。
03. 将冻存管转移到生物安全柜中，并用 70% 乙醇擦拭表面。
04. 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。
05. 在室温下以 125g 离心 5-7 分钟，弃上清。
06. 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。
07. 将细胞转移到 37℃,5% CO2 的培养箱中培养。
08. 参考传代比例：1/2 到 1/4 传代，2-3 天长满。 -
01. 取少量细胞悬液进行细胞计数及活力检测，当细胞密度达到1.5x10^6 cells/mL时，可进行细胞传代。
02. 将培养基从 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。03. 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。
04. 取足量细胞加入盛有新鲜培养基的培养瓶中，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。将细胞密度维持在7x10^5 cells/mL。
05. 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% CO2 培养箱中。
-
01. 准备冻存液，并提前预冷。
02. 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。
03. 将细胞收集到无菌的锥形离心管中并计数细胞。
04. 在室温下以250×g离心细胞5分钟，并小心吸出培养基。
05. 将细胞以至少2x106细胞/mL的密度重悬于冻存液中。
06. 吹打均匀后按照每管 1mL 的量分装至冻存管。
07. 将细胞放在程序降温盒中，在 -80℃冰箱中冷冻。
08. 后续将细胞转移到液氮罐中，以便长期储存。 - 抗体验证中
产品类型： 基因敲除细胞池
细胞系信息
Gene Symbol
ZDHHC3
NCBI Gene ID
51304
Ensembl ID
ENSG00000163812
Uniprot ID
Q9NYG2-2
筛选标记
N/A
宿主细胞 / 类型
K562/人慢性髓原白血病细胞
规格
1×10^6 cells/ 冻存管
生长培养基
RPMI-1640+10% FBS+1% P/S
生长特性
悬浮细胞
培养条件
37℃,5% CO2 的培养箱，1/2 到 1/4 传代
倍增时间
~24-36 hours
参考换液频率
2~3次/周
支原体检测结果
阴性
敲除验证
敲除效率（Sanger测序）
>70%
蛋白质组验证结果
N/A
抗体货号
添加中
抗体验证结果
细胞系说明
目标基因介绍
Golgi-localized palmitoyltransferase that catalyzes the addition of palmitate onto various protein substrates (PubMed:19001095, PubMed:21926431, PubMed:22240897, PubMed:23034182, PubMed:22314500). Has no stringent fatty acid selectivity and in addition to palmitate can also transfer onto target proteins myristate from tetradecanoyl-CoA and stearate from octadecanoyl-CoA (By similarity). Plays an important role in G protein-coupled receptor signaling pathways involving GNAQ and potentially other heterotrimeric G proteins by regulating their dynamic association with the plasma membrane (PubMed:19001095). Palmitoylates ITGA6 and ITGB4, thereby regulating the alpha-6/beta-4 integrin localization, expression and function in cell adhesion to laminin (PubMed:22314500). Plays a role in the TRAIL-activated apoptotic signaling pathway most probably through the palmitoylation and localization to the plasma membrane of TNFRSF10A (PubMed:22240897). In the brain, by palmitoylating the gamma subunit GABRG2 of GABA(A) receptors and regulating their postsynaptic accumulation, plays a role in synaptic GABAergic inhibitory function and GABAergic innervation. Palmitoylates the neuronal protein GAP43 which is also involved in the formation of GABAergic synapses. Palmitoylates NCDN thereby regulating its association with endosome membranes. Probably palmitoylates PRCD and is involved in its proper localization within the photoreceptor. Could mediate the palmitoylation of NCAM1 and regulate neurite outgrowth. Could palmitoylate DNAJC5 and regulate its localization to Golgi membranes. Also constitutively palmitoylates DLG4. May also palmitoylate SNAP25. Could palmitoylate the glutamate receptors GRIA1 and GRIA2 but this has not been confirmed in vivo (By similarity). Could also palmitoylate the D(2) dopamine receptor DRD2 (PubMed:26535572).||May also function as a calcium transporter.
细胞开发路径
采用CRISPR-RNP方法生成稳定KO Cell Pool；Sanger 测序结果显示KO Cell Pool敲除效率>70%。
应用
高敲除效率的基因敲除细胞池（KO Cell Pool），特别适用于初步功能分析、复杂疾病模型的开发、精准药物筛选以及广泛的基因发现研究。KO pool能够无需繁琐的单克隆挑选过程，直接应用于多种类型的测定和分析，大幅提升实验效率。
细胞培养说明
细胞复苏
01. 在 37℃水浴中预热完全培养基。
02. 将冻存管在 37℃水浴中解冻 1-2 分钟。
03. 将冻存管转移到生物安全柜中，并用 70% 乙醇擦拭表面。
04. 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。
05. 在室温下以 125g 离心 5-7 分钟，弃上清。
06. 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。
07. 将细胞转移到 37℃,5% CO2 的培养箱中培养。
08. 参考传代比例：1/2 到 1/4 传代，2-3 天长满。
细胞传代
01. 取少量细胞悬液进行细胞计数及活力检测，当细胞密度达到1.5x10^6 cells/mL时，可进行细胞传代。
02. 将培养基从 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。
03. 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。
04. 取足量细胞加入盛有新鲜培养基的培养瓶中，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。将细胞密度维持在7x10^5 cells/mL。
05. 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% CO2 培养箱中。
细胞冻存
01. 准备冻存液，并提前预冷。
02. 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。
03. 将细胞收集到无菌的锥形离心管中并计数细胞。
04. 在室温下以250×g离心细胞5分钟，并小心吸出培养基。
05. 将细胞以至少2x106细胞/mL的密度重悬于冻存液中。
06. 吹打均匀后按照每管 1mL 的量分装至冻存管。
07. 将细胞放在程序降温盒中，在 -80℃冰箱中冷冻。
08. 后续将细胞转移到液氮罐中，以便长期储存。