CCND1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available.
Images
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Western blot
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.
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
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Western blot
- Disease
Carcinoma
- Concentration
Properties
- Gene name
CCND1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Western blot
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 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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- Culture medium
F-12K + 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
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (Human wild-type A549 cell line ab288558). 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 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.
3 product images
Western blot - Human CCND1 knockout A549 cell line (ab286759)
All lanes: Western blot - Anti-Cyclin D1 antibody [EPR2241] - C-terminal (Anti-Cyclin D1 antibody [EPR2241] - C-terminal ab134175) at 1/10000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: CCND1 knockout A549 cell lysate at 20 µg
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: K562 cell lysate at 20 µg
Western blot - Human CCND1 knockout A549 cell line (ab286759)
False colour image of Western blot: Anti-Cyclin D1 antibody [SP4] staining at 1/25 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 D1 antibody [SP4] ab16663 was shown to bind specifically to Cyclin D1. A band was observed at 35 kDa in wild-type A549 cell lysates with no signal observed at this size in ccnd1 knockout cell line ab286759 (knockout cell lysate ab300213). To generate this image, wild-type and ccnd1 knockout A549 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Cyclin D1 antibody [SP4] (Anti-Cyclin D1 antibody [SP4] ab16663) at 1/25 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: K562 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa
Western blot - Human CCND1 knockout A549 cell line (ab286759)
False colour image of Western blot: Anti-Cyclin D1 antibody - C-terminal staining at 1/25 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 D1 antibody - C-terminal ab185241 was shown to bind specifically to Cyclin D1. A band was observed at 35 kDa in wild-type A549 cell lysates with no signal observed at this size in ccnd1 knockout cell line ab286759 (knockout cell lysate ab300213). To generate this image, wild-type and ccnd1 knockout A549 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Cyclin D1 antibody - C-terminal (Anti-Cyclin D1 antibody - C-terminal ab185241) at 1/25 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 3: SH-SY5Y cell lysate at 20 µg
Lane 4: K562 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa
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