人ATRX knockout A549 cell line
Human ATRX knockout A549 cell line
- Advanced Validation
- 了解详情
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ATRX KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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
X-linked nuclear protein, XH2, Transcriptional regulator ATRX, Znf-HX, XNP, ATP-dependent helicase ATRX, X-linked helicase II, ATRX, RAD54L
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human ATRX knockout A549 cell line (AB287232)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ATRX KO A549 (ATRX knockout human lung carcinoma epithelial cell), ab287232; Wildtype A549 (human lung carcinoma epithelial cell) cells labelling ATRX with ab325390 at 1/500 (1.044 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing nuclear staining in wildtype A549 cells (shown in green), showing no staining in ATRX knockout A549 cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
- WB
Lab
Western blot - Human ATRX knockout A549 cell line (AB287232)
Anti-ATRX antibody staining at 0.4 ug/ml; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution. In Western blot, anti-ATRX antibody was shown to bind specifically to ATRX. A band was observed at 140/190/400 kDa in wild-type A549 cell lysates with no signal observed at this size in ATRX knockout cell line ab287232 (knockout cell lysate ab290802). To generate this image, wild-type and ATRX 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 3 % 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 before development with Optiblot (ECL reagent ab133456) and imaged with 3 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) at 1/50000 dilution and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Anti-ATRX antibody at 0.4 µg/mL
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
Western blot - Human ATRX knockout A549 cell line (ab287232) at 20 µg
Lane 3:
U-2 OS cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Lane 5:
A549 cell lysate at 20 µg
false
搭配使用产品
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.
Recommended control: Human wild-type A549 cell line (ab288558). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
ATRX functions by forming part of the ATRX-DAXX complex. This complex plays a vital role in the deposition of the histone variant H3.3 into heterochromatin regions affecting the structure and function of these genomic segments. ATRX through the ATRX-DAXX complex also maintains telomere integrity by preventing uncontrolled telomere elongation. As such ATRX is important in cellular processes like DNA replication repair and mitotic progression emphasizing its role in maintaining genetic fidelity across cell divisions.
Pathways
ATRX is an important component in chromatin remodeling and telomere maintenance pathways. It interacts closely with the DAXX protein within these pathways affecting the histone exchange mechanisms. ATRX also engages with other proteins such as p53 integrating its function into the DNA damage repair pathway. Through these pathways ATRX ensures proper chromatin dynamics and participates in mechanisms guarding genomic stability.
质量控制
STR 分析
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
Abcam Product Promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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