人PCK1 knockout A549 cell line
Human PCK1 knockout A549 cell line
- Advanced Validation
- 了解详情
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PCK1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
查看别名
GTP, PCK1, PCKGC_HUMAN, PEP carboxykinase, PEPCK-C, PEPCK1, Phosphoenolpyruvate carboxykinase, Phosphoenolpyruvate carboxykinase 1, Phosphoenolpyruvate carboxykinase 1 (soluble), Phosphoenolpyruvate carboxykinase, cytosolic, Phosphoenolpyruvate carboxykinase, cytosolic [GTP], Phosphoenolpyruvate carboxylase, Phosphopyruvate carboxylase, cytosolic [GTP]
- WB
Lab
Western blot - Human PCK1 knockout A549 cell line (AB301135)
Western blot : Anti-PCK1 antibody [EPR6938] (ab133603) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133603 was shown to bind specifically to PCK1. A band was observed at 67 kDa in wild-type A549 cell lysates with no signal observed at this size in PCK1 knockout cell line. To generate this image, wild-type and PCK1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-PCK1/PEPC antibody [EPR6938] (<a href='/products/primary-antibodies/pck1-pepc-antibody-epr6938-ab133603'>ab133603</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
PCK1 knockout A549 cell lysate at 20 µg
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HEK-293 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 67 kDa
false
- NGS
Lab
Next Generation Sequencing - Human PCK1 knockout A549 cell line (AB301135)
1 bp insertion and 35 bp deletion after Met 138 of WT protein
反应性数据
产品详情
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
运输条件
推荐的短期储存条件
推荐的长期储存条件
处理步骤
初始处理指南
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
传代培养指南
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not allow the cell density to exceed 7x104 cells/cm2.
培养基
F-12K + 10% FBS
低温储藏试剂
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
PCK1 is essential for maintaining glucose homeostasis in the body. Although not part of a larger protein complex its function directly impacts the tricarboxylic acid (TCA) cycle by providing substrates for glucose synthesis. By orchestrating the conversion of oxaloacetate to PEP PCK1 contributes significantly to reducing dependency on dietary carbohydrates particularly during fasting or intensive physical activity.
Pathways
PCK1 integrates critical metabolic processes. It mainly influences the gluconeogenesis and glycolysis pathways. In the gluconeogenesis pathway PCK1 coordinates with enzymes such as glucose-6-phosphatase to facilitate the generation of glucose from lactate and amino acids. Additionally its role in glycolysis intersects with enzymes like pyruvate kinase managing energy production and consumption.
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
搭配使用产品
Abcam Product Promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com