SS18 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and 2 bp insertion in exon 4.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and 2 bp insertion in exon 4
Alternative names
MGC116875, Protein SSXT, Protein SYT, SS18, SSXT/SSX4v fusion, SSXT_HUMAN, SYT, SYT SSX1, SYT SSX2, SYT/SSX4v fusion protein, Synovial sarcoma translocated to X chromosome, Synovial sarcoma translocated to X chromosome protein, Synovial sarcoma translocation chromosome 18
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SS18 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and 2 bp insertion in exon 4.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and 2 bp insertion in exon 4
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- SS18
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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 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.- Subculture guidelines
- 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.
- Culture medium
- DMEM (High Glucose) + 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
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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:
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4 product images
Western blot - Human SS18 knockout HeLa cell line (ab265493)
False colour image of Western blot: Anti-SS18 antibody [EPR23636-98] staining at 1/1000 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-SS18 antibody [EPR23636-98] ab254405 was shown to bind specifically to SS18. A band was observed at 51/56 kDa in wild-type HeLa cell lysates with no signal observed at this size in SS18 knockout cell line ab265493 (knockout cell lysate Human SS18 knockout HeLa cell lysate ab259156). To generate this image wild-type and SS18 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-SS18 antibody [EPR23636-98] (Anti-SS18 antibody [EPR23636-98] ab254405) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: SS18 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human SS18 knockout HeLa cell line (ab265493)
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 51 kDa, 56 kDa
Western blot - Human SS18 knockout HeLa cell line (ab265493)
False colour image of Western blot: Anti-SS18 antibody staining at 1/500 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-SS18 antibody ab272904 was shown to bind specifically to SS18. A band was observed at 51/60 kDa in wild-type HeLa cell lysates with no signal observed at this size in SS18 knockout cell line ab265493 (knockout cell lysate Human SS18 knockout HeLa cell lysate ab259156). To generate this image wild-type and SS18 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-SS18 antibody (Anti-SS18 antibody ab272904) at 1/500 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: SS18 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human SS18 knockout HeLa cell line (ab265493)
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 51 kDa, 60 kDa
Sanger Sequencing - Human SS18 knockout HeLa cell line (ab265493)
Allele-2: 2 bp insertion in exon 4.
Sanger Sequencing - Human SS18 knockout HeLa cell line (ab265493)
Allele-1: 1 bp insertion in exon 4.
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