CCND2 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 8 bp insertion in exon 1.
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- Journal of oncology 2022:54560162022miR-187/PDLIM1 Gets Involved in Gastric Cancer Progression and Cisplatin Sensitivity of Cisplatin by Mediating the Hippo-YAP Signaling Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYeru Tan,Yuehua Li,Hongbo Zhu,Xiaoping Wu,Kai Mei,Pian Li,Qiao YangPubMed 36164345
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 8 bp insertion in exon 1
Alternative names
CCND2_HUMAN, CyclinD2, G1/S-specific cyclin-D2, KIAK0002, MGC102758, MPPH3
Recommended products
CCND2 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 8 bp insertion in exon 1.
Alternative names
CCND2_HUMAN, CyclinD2, G1/S-specific cyclin-D2, KIAK0002, MGC102758, MPPH3
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 8 bp insertion in exon 1
- Concentration
Properties
- Gene name
CCND2
- 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 long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
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.
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.
3 product images
Western blot - Human CCND2 (Cyclin D2) knockout HEK-293T cell line (ab267318)
All lanes: Western blot - Anti-Cyclin D2 antibody [EPR19659] (Anti-Cyclin D2 antibody [EPR19659] ab207604) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human CCND2 (Cyclin D2) knockout HEK-293T cell lysate (Human CCND2 (Cyclin D2) knockout HEK-293T cell lysate ab257875) at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: U-2 OS cell lysate at 20 µg
SecondaryLanes 1 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 33 kDa
Observed band size: 33 kDa
False colour image of Western blot: Anti-Cyclin D2 antibody [EPR19659] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Cyclin D2 antibody [EPR19659] ab207604 was shown to bind specifically to Cyclin D2. A band was observed at 33 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in Ccnd2 knockout cell line ab267318 (knockout cell lysate Human CCND2 (Cyclin D2) knockout HEK-293T cell lysate ab257875). To generate this image, wild-type and Ccnd2 knockout HEK-293T 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 °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 (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.
Sanger Sequencing - Human CCND2 (Cyclin D2) knockout HEK-293T cell line (ab267318)
Allele-1: 2 bp deletion in exon1
Sanger Sequencing - Human CCND2 (Cyclin D2) knockout HEK-293T cell line (ab267318)
Allele-2: 8 bp insertion in exon 1.
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