人TMPRSS2 knockout Caco-2 cell line (ab273737)
概述
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产品名称
人TMPRSS2 knockout Caco-2 cell line
参阅全部 TMPRSS2 细胞裂解液 -
Parental Cell Line
Caco 2 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 77bp deletion in exon 3 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Biosafety level
1 -
常规说明
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: EMEM + 20% 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 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 1x104 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 (approx 8x104-1x105 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
Colon -
Cell type
epithelial -
Disease
Adenocarcinoma -
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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组织特异性
Expressed strongly in small intestine. Also expressed in prostate, colon, stomach and salivary gland. -
序列相似性
Belongs to the peptidase S1 family.
Contains 1 LDL-receptor class A domain.
Contains 1 peptidase S1 domain.
Contains 1 SRCR domain. -
细胞定位
Cell membrane and Secreted. Activated by cleavage and secreted. - Information by UniProt
图片
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All lanes : Anti-TMPRSS2 antibody [EPR3862] (ab109131) at 1/2000 dilution
Lane 1 : Wild-type LNCaP cell lysate
Lane 2 : TMPRSS2 knockout LNCaP cell lysate
Lane 3 : Wild-type Caco-2 cell lysate
Lane 4 : TMPRSS2 knockout Caco-2 cell lysate
Lane 5 : PC-3 cell lysate
Lane 6 : DU 145 cell lysate
Lane 7 : Human Prostate cell lysate
Lane 8 : Human Colon cell lysate
Lane 9 : Human Lung cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 55,25 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-TMPRSS2 antibody [EPR3862] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109131 was shown to bind specifically to TMPRSS2. A band was observed at 55, 25 kDa in wild-type LNCaP and at 25 kDa in Caco-2 cell lysates with no signal observed at this size in TMPRSS2 knockout LNCaP cell line ab273745 (knockout LNCaP cell lysate ab275499) and TMPRSS2 knockout Caco-2 cell line ab273737 (knockout Caco-2 cell lysate ab277340). To generate this image, wild-type and TMPRSS2 knockout cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Allele-1: 77bp deletion in exon 3
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)
ab273737 尚未被引用在任何文献中。