人CD9 knockout A549 cell line (ab261878)
概述
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产品名称
人CD9 knockout A549 cell line
参阅全部 CD9 细胞裂解液 -
Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 7 bp deletion; 1 bp deletion; Frameshift = 100% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
经测试应用
适用于: Next Generation Sequencing, WBmore details -
Biosafety level
1 -
常规说明
Recommended control: Human wild-type A549 cell line (ab259777). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM:Hams F12 + 5% FBS
Initial handling guidelines: 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x103-1x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.Subculture guidelines:
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 6x104 cells/cm2 is recommended.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not exceed 7x104 cells/cm2.
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.
We will provide viable cells that proliferate on revival.
性能
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Lung -
Cell type
epithelial -
Disease
Carcinoma -
Gender
Male -
Mycoplasma free
Yes -
存放说明
Shipped on Dry Ice. Store in liquid nitrogen. -
存储溶液
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
研究领域
靶标
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功能
Involved in platelet activation and aggregation. Regulates paranodal junction formation. Involved in cell adhesion, cell motility and tumor metastasis. Required for sperm-egg fusion. -
组织特异性
Expressed by a variety of hematopoietic and epithelial cells. -
序列相似性
Belongs to the tetraspanin (TM4SF) family. -
翻译后修饰
Protein exists in three forms with molecular masses between 22 and 27 kDa, and is known to carry covalently linked fatty acids. -
细胞定位
Membrane. - Information by UniProt
相关产品
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KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab261878于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Next Generation Sequencing |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration.
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说明 |
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Next Generation Sequencing
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. |
图片
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7 bp deletion (allele 1) and 1 bp deletion (allele 2) after Ile30 of the WT protein
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All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : CD9 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Performed under reducing conditions.Lanes 1 - 3: Merged signal (red and green). Green - ab92726 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab92726 was shown to recognize CD9 in wild-type A549 cells as signal was lost at the expected MW in CD9 knockout cell line ab261878 (knockout cell lysate ab261687). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and CD9 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Ab92726 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C 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.
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Knockout achieved by CRISPR/Cas9; X = 7 bp deletion, 1 bp deletion; Frameshift = 100%
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (0)
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