CRBN KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Sanger Sequencing, Western blot
Alternative names
2610203G15Rik, 2900045O07Rik, AF229032, AW108261, CRBN_HUMAN, Cereblon, DKFZp781K0715, MGC27358, MRT2A, OTTHUMP00000209555, Protein cereblon, Protein x 0001, piL
Recommended products
CRBN KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Sanger Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- CRBN
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Sanger 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 short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Recommended control: 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: McCoY5a + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
• A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
• Cells should be passaged when they have achieved 80-90% confluence.
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
CRBN also known as cereblon is a protein involved in the regulation of protein homeostasis. It has a molecular weight of approximately 51 kDa. CRBN is expressed in various tissues including the brain kidney and muscle. It functions as a substrate receptor for the CRL4-CRBN E3 ubiquitin ligase complex recognizing proteins for ubiquitination and degradation. The chemical structure of thalidomide shows binding affinity to CRBN influencing its function.
- Biological function summary
Cereblon plays a role in the regulation of protein stability and degradation. It forms part of the CRL4-CRBN complex which tags specific proteins for ubiquitination and subsequent proteasomal degradation. This function is critical for maintaining cellular protein balance. Additionally CRBN modulates immune response and has immunomodulator properties influencing cell-mediated responses.
- Pathways
CRBN participates in the ubiquitin-proteasome pathway which regulates protein turnover and quality control within cells. Moreover CRBN is involved in immune signaling pathways partly by modulating the expression of various cytokines and immune-related proteins. It interacts with proteins such as DDB1 and CUL4 within these pathways impacting cellular responses to stress and immune challenges.
- Associated diseases and disorders
CRBN is associated with multiple myeloma and intellectual disability. Cereblon acts as a target for drugs like thalidomide and lenalidomide which are used in the treatment of multiple myeloma by altering ubiquitination processes. In intellectual disability alterations in CRBN function or expression can disrupt neuronal development and signaling. CRBN interacts with proteins like IKZF1 and IKZF3 within these contexts affecting their roles in disease progression and treatment responses.
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 CRBN knockout HCT116 cell line (ab289186)
Anti-CRBN antibody [4D6] (Anti-CRBN antibody [4D6] ab244223) staining at 2 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CRBN antibody [4D6] ab244223 was shown to bind specifically to CRBN. A band was observed at 51 kDa in wild-type cell lysates with no signal observed at this size in CRBN knockout cell lines. To generate this image, wild-type and CRBN knockout cell lysates were analysed: CRBN knockout A549 cell line (ab288959), CRBN knockout HCT 116 cell line (ab288959) and CRBN knockout MOLT-4. 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-CRBN antibody [4D6] (Anti-CRBN antibody [4D6] ab244223) at 2 µg/mL
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: Western blot - Human CRBN knockout A549 cell line (ab288959)
Lane 2: CRBN knockout A549 cell lysate at 20 µg
Lane 3: Wild-type HCT 116 cell lysate at 20 µg
Lane 4: Western blot - Human CRBN knockout HCT116 cell line (ab289186)
Lane 4: CRBN knockout HCT 116 cell lysate at 20 µg
Lane 5: Wild-type MOLT-4 cell lysate at 20 µg
Lane 6: CRBN knockout MOLT-4 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.
Predicted band size: 51 kDa
Observed band size: 51 kDa
Next Generation Sequencing - Human CRBN knockout HCT116 cell line (ab289186)
83 bp deletion after Ser76 of the WT protein.
Sanger Sequencing - Human CRBN knockout HCT116 cell line (ab289186)
83bp deletion after Ser77 of the WT protein.
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com