人ITGB4 knockout A549 cell line
Human ITGB4 knockout A549 cell line
- Advanced Validation
- 了解详情
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ITGB4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
查看别名
CD 104, CD104 antigen, ITB4_HUMAN, ITG B4, Integrin beta 4 subunit, Integrin beta-4, gp150
- WB
Lab
Western blot - Human ITGB4 knockout A549 cell line (AB301019)
Western blot : Anti-ITGB4 antibody [EPR8558(2)] (ab168386) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab168386 was shown to bind specifically to ITGB4. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in ITGB4 knockout cell line. To generate this image, wild-type and ITGB4 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 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-Integrin beta 4 antibody [EPR8558(2)] (<a href='/products/primary-antibodies/integrin-beta-4-antibody-epr85582-ab168386'>ab168386</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 cell lysate at 40 µg
Lane 2:
ITGB4 knockout A549 cell lysate at 40 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: 200 kDa
false
- WB
Lab
Western blot - Human ITGB4 knockout A549 cell line (AB301019)
Western blot : Anti-ITGB4 antibody [EPR17517] (ab182120) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab182120 was shown to bind specifically to ITGB4. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in ITGB4 knockout cell line. To generate this image, wild-type and ITGB4 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 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-Integrin beta 4 antibody [EPR17517] (<a href='/products/primary-antibodies/integrin-beta-4-antibody-epr17517-ab182120'>ab182120</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 40 µg
Lane 2:
ITGB4 knockout A549 cell lysate at 40 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: 200 kDa
false
- NGS
Lab
Next Generation Sequencing - Human ITGB4 knockout A549 cell line (AB301019)
14 bp deletion after His 267 and 131 bp deletion after Val 282 (allele 1); 139 bp deletion after His 267 (allele 2); 9 bp deletion and 1 bp insertion after His 267 and 2 bp deletion after Cys 284 (allele 3); 3 bp insertion after His 267 (allele 4)
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
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
Integrin beta 4 is important for anchoring epithelial cells to the basement membrane stabilizing cell layers in tissues. It interacts with intermediate filaments through specific cytoplasmic proteins. The integrin beta 4 complex ensures structural integrity essential for maintaining the skin and mucosal barriers. Integrations with proteins like M126 are essential for strengthening these interactions safeguarding cellular stability and promoting wound healing.
Pathways
Integrin beta 4 is embedded in key signaling pathways that influence cell adhesion and migration. It participates prominently in the PI3K/AKT and RAS pathway collaborations facilitating signal transduction essential for cell growth and survival. Through these pathways integrin beta 4 interacts with proteins like IS200BPIRG1A linking extracellular cues to internal cellular responses.
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
Complementary Products
Primary Antibodies
AB182120
Anti-Integrin beta 4 antibody [EPR17517]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-Integrin beta 4 antibody [EPR17517] (AB182120)](https://content.abcam.com/products/images/integrin-beta-4-antibody-epr17517-ab182120--western-blot-img436368.jpg)
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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