Human VCAM1 knockout A549 cell line
Human VCAM1 knockout A549 cell line
- Advanced Validation
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VCAM1 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
別名を表示する
CD106, CD106 Antigen, INCAM-100, MGC99561, VCAM1_HUMAN, Vascular Cell Adhesion Molecule 1, Vascular cell adhesion protein 1
- Flow Cyt
Lab
Flow Cytometry - Human VCAM1 knockout A549 cell line (AB273758)
Flow cytometry overlay histogram showing wild-type A549 (green line) and VCAM1 knockout A549 cells (red line, ab273758), treated with 10 ng/ml TNF-alpha for 16 h (left) and untreated (right), stained with ab103173. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab103173) (1x106 in 100μl at 0.2μg/ml) for 30 min at 4°C.
Isotype control antibody mouse IgG1κ Allophycocyanin was used at the same concentration and conditions as the primary antibody (wild-type A549 - black line VCAM knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.
- WB
Lab
Western blot - Human VCAM1 knockout A549 cell line (AB273758)
Lanes 1 - 6 : Merged signal (red and green). Green - ab134047 observed at 105 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab134047 was shown to react with VCAM1 in treated wild-type A549 cells in Western blot with loss of signal observed in treated VCAM1 knockout cell line ab273758 (knockout cell lysate ab275504). Wild-type A549 and VCAM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab134047 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-VCAM1 antibody [EPR5047] (<a href='/products/primary-antibodies/vcam1-antibody-epr5047-ab134047'>ab134047</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 cell lysate at 30 µg
Lane 2:
Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg
Lane 2:
Western blot - Human VCAM1 knockout A549 cell line (ab273758)
Lane 3:
VCAM1 knockout A549 cell lysate at 30 µg
Lane 4:
VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg
Lane 5:
HUVEC cell lysate at 30 µg
Lane 6:
HUVEC TNF-a treated (16 ng/mL, 16h) cell lysate at 30 µg
Predicted band size: 81 kDa
Observed band size: 105 kDa
false
- WB
Lab
Western blot - Human VCAM1 knockout A549 cell line (AB273758)
ab174279 was shown to react with VCAM1 in treated wild-type A549 cells in western blot. Loss of signal was observed when treated VCAM1 knockout cell line ab273758 (knockout cell lysate ab275504) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab174279 overnight at 4 ° at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.
All lanes:
Western blot - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 30 µg
Lane 2:
Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg
Lane 3:
VCAM1 knockout A549 cell lysate at 30 µg
Lanes 3 - 4:
Western blot - Human VCAM1 knockout A549 cell line (ab273758)
Lane 4:
VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg
Predicted band size: 81 kDa
Observed band size: 105 kDa
false
Exposure time: 20s
- Sanger seq
Lab
Sanger Sequencing - Human VCAM1 knockout A549 cell line (AB273758)
Allele-1 : 4 bp deletion in exon 2
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
接合型
出荷温度
短期保存温度
長期保存温度
取り扱い方法
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.
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.
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.
補足情報
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Adhesion molecules like VCAM1 participate in immune response by recruiting leukocytes to sites of inflammation. VCAM1 helps guide immune cells to directed locations where they perform defense activities. Although VCAM1 is not part of any known large protein complex its interaction with integrins facilitates the migration of immune cells across the endothelium and into tissue.
Pathways
VCAM1 contributes significantly to the leukocyte extravasation process in inflammatory pathways. It plays a role in both the cytokine-cytokine receptor interaction pathway and the NF-kB signaling pathway which are essential for immune response and cell signaling. Through these pathways it interacts with proteins like ICAM1 and various chemokines helping to coordinate the immune cell movement through vascular tissue barriers.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
組合せ製品
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