Human NT5E knockout A-431 cell line
Human NT5E knockout A-431 cell line
- Advanced Validation
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NT5E KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 5 bp deletion, 2 bp deletion; Frameshift = 99.2%. 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
Ecto-5'-nucleotidase, NT5, Thymidylate 5'-phosphatase, CD73, NTE, 5'-NT, 5'-deoxynucleotidase, IMP-specific 5'-nucleotidase, NT5E, 5'-nucleotidase
- WB
Lab
Western blot - Human NT5E knockout A-431 cell line (AB261895)
Lanes 1 - 3 : Merged signal (red and green). Green - ab202122 observed at 63 kDa. Red - loading control ab181602 observed at 37 kDa.
ab202122 was shown to recognize NT5E in wild-type A-431 cells as signal was lost at the expected MW in NT5E knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. ab202122 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4° at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-CD73 antibody [4G6E3] (<a href='/products/primary-antibodies/cd73-antibody-4g6e3-ab202122'>ab202122</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 whole cell lysate at 20 µg
Lane 2:
NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human NT5E knockout A-431 cell line (ab261895)
Lane 3:
A375 (Human malignant melanoma cell line) whole cell lysate at 20 µg
Predicted band size: 63 kDa
false
- WB
Lab
Western blot - Human NT5E knockout A-431 cell line (AB261895)
Lanes 1 - 3 : Merged signal (red and green). Green - ab133582 observed at 63 kDa. Red - loading control ab8245 observed at 37 kDa.
ab133582 was shown to recognize NT5E in wild-type A-431 cells as signal was lost at the expected MW in NT5E knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab133582 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4° at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-CD73 antibody [EPR6114] (<a href='/products/primary-antibodies/cd73-antibody-epr6114-ab133582'>ab133582</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human NT5E knockout A-431 cell line (ab261895)
Lane 3:
A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg
Predicted band size: 105 kDa,143 kDa,24 kDa,35 kDa,41 kDa,44 kDa,45 kDa,53 kDa,58 kDa,61 kDa,62 kDa,63 kDa,70 kDa,88 kDa
Observed band size: 104 kDa,18 kDa,55 kDa,58 kDa
false
- NGS
Lab
Next Generation Sequencing - Human NT5E knockout A-431 cell line (AB261895)
x = 5 bp deletion, 2 bp deletion
- NGS
Supplier Data
Next Generation Sequencing - Human NT5E knockout A-431 cell line (AB261895)
2 bp deletion after Asp110 (allele 1) and 5 bp deletion after Tyr109 (allele 2) of the WT protein
組合せ製品
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
短期保存温度
長期保存温度
補足情報
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
CD73 participates in the regulation of purinergic signaling through adenosine production. It acts in cooperation with other cell surface enzymes such as CD39 forming a functional complex that handles ATP breakdown to adenosine. This enzymatic activity modulates immune responses tissue protection and inflammation control. In addition to immune-related functions CD73 supports the maintenance of vascular integrity and proper cellular adhesion which is important in different physiological and pathological contexts.
Pathways
CD73 plays an important part in the adenosinergic pathway governing the conversion of AMP into adenosine. This activity impacts the signaling and function of the adenosine receptors on immune and non-immune cells. Another important pathway is the angiogenesis process where CD73 contributes by affecting endothelial cell migration and blood vessel development. Relationships with key proteins such as adenosine A2 receptors help facilitate these critical pathway activities.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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.
培養培地
DMEM (High Glucose) + 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.
ターゲットの情報
Abcam product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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