CCND1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
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Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1
Alternative names
AI327039, B cell CLL/lymphoma 1, B cell leukemia 1, B-cell lymphoma 1 protein, BCL-1, BCL-1 oncogene, CCND1/FSTL3 fusion gene, CCND1/FSTL3 fusion gene, included, CCND1/IGHG1 fusion gene, CCND1/IGHG1 fusion gene, included, CCND1/IGLC1 fusion gene, CCND1/IGLC1 fusion gene, included, CCND1/PTH fusion gene, CCND1/PTH fusion gene, included, CCND1_HUMAN, CD1, Cyl 1, D11S287E, G1/S-specific cyclin-D1, PRAD1, PRAD1 oncogene, Parathyroid adenomatosis 1, U21B31
Recommended products
CCND1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
Alternative names
AI327039, B cell CLL/lymphoma 1, B cell leukemia 1, B-cell lymphoma 1 protein, BCL-1, BCL-1 oncogene, CCND1/FSTL3 fusion gene, CCND1/FSTL3 fusion gene, included, CCND1/IGHG1 fusion gene, CCND1/IGHG1 fusion gene, included, CCND1/IGLC1 fusion gene, CCND1/IGLC1 fusion gene, included, CCND1/PTH fusion gene, CCND1/PTH fusion gene, included, CCND1_HUMAN, CD1, Cyl 1, D11S287E, G1/S-specific cyclin-D1, PRAD1, PRAD1 oncogene, Parathyroid adenomatosis 1, U21B31
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, Homozygous: Insertion of the selection cassette in exon 1
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
CCND1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
- Zygosity
Homozygous
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 HeLa cell line (Human wild-type HeLa cell line ab255448). 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 CCND1 (Cyclin D1) knockout HeLa cell line (ab261760)
Lane 1: Wild-type HeLa cell lysate (20μg)
Lane 2: CCND1 knockout HeLa cell lysate (20μg)
Lanes 1- 2: Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.
ab40754 Anti-Cyclin D1 antibody [EP272Y] was shown to specifically react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261760 (knockout cell lysate Human CCND1 (Cyclin D1) knockout HeLa cell lysate ab256864) was used. Wild-type and Cyclin D1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40754 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.All lanes: Western blot - Anti-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CCND1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 36 kDa
Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab261760)
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cyclin D1 antibody [SP4] ab16663 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-Cyclin D1 antibody [SP4] ab16663 was shown to react with Cyclin D1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261760 (knockout cell lysate Human CCND1 (Cyclin D1) knockout HeLa cell lysate ab256864) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Cyclin D1 antibody [SP4] ab16663 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) overnight at 4° at a 1 in 200 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Cyclin D1 antibody [SP4] (Anti-Cyclin D1 antibody [SP4] ab16663) at 1/200 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CCND1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 36 kDa
Sanger Sequencing - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab261760)
Homozygous: Insertion of the selection cassette in exon 1.
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