Human VIL1 Knockout A549 cell line
Human VIL1 Knockout A549 cell line
- Advanced Validation
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- Sanger seq
Lab
Sanger Sequencing - Human VIL1 Knockout A549 cell line (AB324222)
256 bp deletion after Val148 (edit 1); 190 bp deletion and 1 bp deletion after Val148 (edit 2)
- WB
Lab
Western blot - Human VIL1 Knockout A549 cell line (AB324222)
Western blot : Anti-VIL1 antibody [EPR3491(3)] (ab133510) staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133510 was shown to bind specifically to VIL1. A band was observed at 93 kDa in wild-type A549 cell lysates with no signal observed at this size in VIL1 knockout cell line. To generate this image, wild-type and VIL1 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-Villin antibody [EPR3491(3)] (<a href='/products/primary-antibodies/villin-antibody-epr34913-ab133510'>ab133510</a>) at 1/5000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lanes 2 and 4:
Empty cell lysate at 20 µg
Lane 3:
Western blot - Human VIL1 Knockout A549 cell line (ab324222) at 20 µg
Lane 5:
HepG2 cell lysate at 20 µg
Lane 6:
Jurkat Nuclear 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: 93 kDa
false
Reactivity data
製品の詳細
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.
製品内容
出荷温度及び保存条件
遺伝子名
遺伝子編集のタイプ
遺伝子編集の方法
ノックアウト検証方法
出荷温度
短期保存期間
短期保存温度
長期保存温度
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Viability
>= 60%
初回取り扱いガイドライン
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>
継代培養ガイドライン
- 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.
培養培地
F-12K + 10% FBS
凍結保存培地
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'.
For licensing inquiries, please contact partnerships@abcam.com