CDKN1B KO cell line available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available.
Images
Key facts
- Cell type
HCT116
- Species or organism
Human
- Tissue
Colon
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
Alternative names
AA408329, AI843786, CDKN 1B, CDKN 4, CDN1B_HUMAN, Cdki1b, Cyclin-dependent kinase inhibitor 1B, Cyclin-dependent kinase inhibitor 1B (p27, Kip1), Cyclin-dependent kinase inhibitor p27, Cyclin-dependent kinase inhibitor p27 Kip1, KIP 1, MEN1B, MEN4, OTTHUMP00000195098, OTTHUMP00000195099, P27-like cyclin-dependent kinase inhibitor, p27, p27 Kip1
Recommended products
CDKN1B KO cell line available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available.
Alternative names
AA408329, AI843786, CDKN 1B, CDKN 4, CDN1B_HUMAN, Cdki1b, Cyclin-dependent kinase inhibitor 1B, Cyclin-dependent kinase inhibitor 1B (p27, Kip1), Cyclin-dependent kinase inhibitor p27, Cyclin-dependent kinase inhibitor p27 Kip1, KIP 1, MEN1B, MEN4, OTTHUMP00000195098, OTTHUMP00000195099, P27-like cyclin-dependent kinase inhibitor, p27, p27 Kip1
Key facts
- Cell type
HCT116
- Species or organism
Human
- Tissue
Colon
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Disease
Carcinoma
- Concentration
Properties
- Gene name
CDKN1B
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Next Generation Sequencing, 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.
- Culture medium
McCoY5a + 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 HCT116 cell line (Human wild-type HCT116 cell line ab288559). 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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The protein known as p27 KIP1 also referred to as p27 CDKN1B or KIP1 is a member of the kinase inhibitory protein family. It plays an important role in cell cycle regulation by inhibiting cyclin-dependent kinases (CDKs). p27 KIP1 weighs approximately 27 kDa and can be found in various tissues where it regulates cell proliferation. The protein functions as a suppressor by binding to cyclin-CDK complexes preventing the transition from G1 phase to S phase in the cell division cycle.
- Biological function summary
The function of p27 KIP1 involves its role in controlling cell growth and division. It achieves this by becoming a part of larger protein complexes involving CDKs and cyclins. By directly interacting with these complexes p27 KIP1 modulates cell cycle progression therefore acting as a brake on cellular proliferation. The protein is important in maintaining proper cell cycle checkpoints and preventing uncontrolled cell growth which is essential for normal cellular functioning.
- Pathways
The involvement of p27 KIP1 centers on cell cycle regulation and signaling pathways such as the PI3K/AKT pathway. Its interaction with CDKs and cyclins situates it within the core mechanisms that determine cell division timing. p27 KIP1 operates alongside other proteins like cyclin D and CDK4/6 fitting into the regulatory intricacies of these pathways. Proper functioning of these pathways ensures cellular homeostasis and prevents the development of oncogenic processes.
- Associated diseases and disorders
Dysfunction of p27 KIP1 has links to cancer and neurodegenerative diseases. Low levels or mutations can lead to uncontrolled cell proliferation contributing to the development and progression of cancers such as breast cancer. p27 KIP1's role in neurodegenerative diseases involves its regulation of neuronal cell cycle re-entry with abnormalities potentially exacerbating conditions like Alzheimer's disease. In these contexts its interaction with proteins such as cyclin E and CDK2 becomes particularly relevant in understanding disease mechanisms.
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3 product images
Western blot - Human CDKN1B knockout HCT116 cell line (ab286580)
Western blot: Anti-CDKN1B antibody [Y236] (Anti-p27 KIP 1 antibody [Y236] ab32034) staining at 1/5000 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 magenta. In Western blot, Anti-p27 KIP 1 antibody [Y236] ab32034 was shown to bind specifically to CDKN1B. A band was observed at 30 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CDKN1B knockout cell line. To generate this image, wild-type and CDKN1B knockout HCT 116 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-p27 KIP 1 antibody [Y236] (Anti-p27 KIP 1 antibody [Y236] ab32034) at 1/5000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: CDKN1B knockout HCT 116 cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: CDKN1B knockout HAP1 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.
Western blot - Human CDKN1B knockout HCT116 cell line (ab286580)
Western blot: Anti-CDKN1B antibody [SX53G8] (Anti-p27 KIP 1 antibody [SX53G8] ab193379) staining at 1 ug/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-p27 KIP 1 antibody [SX53G8] ab193379 was shown to bind specifically to CDKN1B. A band was observed at 30 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CDKN1B knockout cell line. To generate this image, wild-type and CDKN1B knockout HCT 116 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-p27 KIP 1 antibody [SX53G8] (Anti-p27 KIP 1 antibody [SX53G8] ab193379) at 1 µg/mL
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: CDKN1B knockout HCT 116 cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: CDKN1B knockout HAP1 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 30 kDa
Next Generation Sequencing - Human CDKN1B knockout HCT116 cell line (ab286580)
103 bp deletion after Val5 (allele 1); 11 bp deletion after Val5, 1 bp insertion downstream (allele 2)
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