TMPRSS2 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,
Homozygous: 79 bp deletion in exon 3.
Images
Key facts
- Cell type
- LNCaP
- Species or organism
- Human
- Tissue
- Prostate
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 79 bp deletion in exon 3
Alternative names
D16Ertd61e, Epitheliasin, FLJ41954, MGC6821, PP9284, PRSS10, Serine protease 10, TMPRSS2, TMPRSS2 ERG FUSION GENE, INCLUDED, TMPRSS2 ETV1 FUSION GENE, INCLUDED, TMPS2_HUMAN, Transmembrane protease serine 2 catalytic chain, Transmembrane protease, serine 2, Transmembrane protease, serine 2, EC 3.4.219
Recommended products
TMPRSS2 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,
Homozygous: 79 bp deletion in exon 3.
Key facts
- Cell type
- LNCaP
- Species or organism
- Human
- Tissue
- Prostate
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 79 bp deletion in exon 3
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- TMPRSS2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Immunocytochemistry, Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
Handling procedures
- 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 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 60-70% confluent.
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.- Subculture guidelines
- Very slow growing. Split once a week and media change once in between splits.
- Cells grow in loosely attached "islands".
- Cells should be passaged when they are 60-70% confluent. Do not allow confluency to exceed this.
- Be careful when washing to not detach cells.
- Use dissociation reagent for a maximum of 1 minute.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 1x104 cells/cm2 is recommended.
- Culture medium
- RPMI + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
5 product images
Western blot - Human TMPRSS2 knockout LNCaP cell line (ab273745)
False colour image of Western blot: Anti-TMPRSS2 antibody [EPR3862] staining at 1/2000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red.
In Western blot Anti-TMPRSS2 antibody [EPR3862] 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 Human TMPRSS2 knockout LNCaP cell lysate ab275499) and TMPRSS2 knockout Caco-2 cell line Human 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 °. 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-TMPRSS2 antibody [EPR3862] (Anti-TMPRSS2 antibody [EPR3862] ab109131) at 1/2000 dilution
Lane 1: Wild-type LNCaP cell lysate at 20 µg
Lane 2: TMPRSS2 knockout LNCaP cell lysate at 20 µg
Lane 2: Western blot - Human TMPRSS2 knockout LNCaP cell line (ab273745)
Lane 3: Wild-type Caco-2 cell lysate at 20 µg
Lane 4: TMPRSS2 knockout Caco-2 cell lysate at 20 µg
Lane 5: PC-3 cell lysate at 20 µg
Lane 6: DU 145 cell lysate at 20 µg
Lane 7: Human Prostate cell lysate at 20 µg
Lane 8: Human Colon cell lysate at 20 µg
Lane 9: Human Lung cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 25 kDa, 55 kDa
Immunocytochemistry/ Immunofluorescence - Human TMPRSS2 knockout LNCaP cell line (ab273745)
Anti-TMPRSS2 antibody [EPR24407-87] ab280567 staining TMPRSS2 in wild-type LnCAP cells (top panel) and TMPRSS2 knockout LnCAP cells (ab273745) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-TMPRSS2 antibody [EPR24407-87] ab280567 at 1μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Sandwich ELISA - Human TMPRSS2 knockout LNCaP cell line (ab273745)
Human TMPRSS2 concentration was interpolated from the TMPRSS2 standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human TMPRSS2 ELISA Kit (Human TMPRSS2 ELISA Kit ab283552). Wild-type and TMPRSS2 knockout LNCaP (ab273745) cells were assessed in duplicate (n=2). Data are represented as the mean and error bars represent standard deviation.
Sanger Sequencing - Human TMPRSS2 knockout LNCaP cell line (ab273745)
79 bp deletion in exon 3
Western blot - Human TMPRSS2 knockout LNCaP cell line (ab273745)
Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBSLysates at 20 µg per lane.
The samples were run on a Bis-Tris gel under reducing conditions.
Western blot: Anti-TMPRSS2 antibody (Anti-TMPRSS2 antibody [P5H9-A3] ab309546) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red.
In Western blot, Anti-TMPRSS2 antibody [P5H9-A3] ab309546 was shown to bind specifically to TMPRSS2. Target of interest was observed at 55 kDa and 25 KDa in wild-type LNCaP cell lysates (lane 1) with no signal observed at this size in TMPRSS2 knockout cell line (lane 2, knockout cell line ab273745). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
The molecular weight observed is consistent with what has been described in the literature (PMID: 35104687).
Exposure time:All lanes: Western blot - Anti-TMPRSS2 antibody [P5H9-A3] (Anti-TMPRSS2 antibody [P5H9-A3] ab309546) at 1/1000 dilution
Lane 1: Wild-type LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: TMPRSS2 knockout LNCaP whole cell lysate at 20 µg
SecondaryLanes 1 - 2: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/200000 dilution
Lanes 1 - 2: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/200000 dilution
Observed band size: 25, 55 kDa
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