DYRK1A KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 12.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 12
Alternative names
DYR1A_HUMAN, DYRK, DYRK 1, DYRKA, Dual specificity YAK 1 related kinase, Dual specificity YAK1-related kinase, Dual specificity tyrosine (Y) phosphorylation regulated kinase 1A, Dual specificity tyrosine phosphorylation regulated kinase, Dual specificity tyrosine phosphorylation regulated kinase 1, Dual specificity tyrosine-phosphorylation-regulated kinase 1A, HP 86, MNB, MNB protein kinase, MNB protein kinase, Serine/threonine-specific, MNB/DYRK protein kinase, MNBH, MRD7, Minibrain (Drosophila) homolog, Minibrain homolog, Minibrain, Drosophila, homolog of, Mnb protein kinase homolog hp86, OTTHUMP00000109090, OTTHUMP00000109091, OTTHUMP00000109094, OTTHUMP00000174799, Protein kinase minibrain homolog, Serine/threonine kinase MNB, hMNB
Recommended products
DYRK1A KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 12.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 12
- Concentration
Properties
- Gene name
- DYRK1A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- 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 short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
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3 product images
Western blot - Human DYRK1A knockout HEK-293T cell line (ab267336)
False colour image of Western blot: Anti-DYRK1A antibody [EPR24132-61] staining at 1/2000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-DYRK1A antibody [EPR24132-61] ab259869 was shown to bind specifically to DYRK1A. A band was observed at 80/85/90 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in DYRK1A CRISPR-Cas9 edited cell line ab267336 (CRISPR-Cas9 edited cell lysate Human DYRK1A knockout HEK-293T cell lysate ab258402). The band observed in the CRISPR-Cas9 edited lysate lane below 80/85/90 kDa is likely to represent a truncated form of DYRK1A. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and DYRK1A CRISPR-Cas9 edited 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 °. 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.
All lanes: Western blot - Anti-DYRK1A antibody [EPR24132-61] (Anti-DYRK1A antibody [EPR24132-61] ab259869) at 1/2000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: DYRK1A CRISPR-Cas9 edited HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human DYRK1A knockout HEK-293T cell line (ab267336)
Lane 3: HT1080 cell lysate at 20 µg
Lane 4: U-251 MG cell lysate at 20 µg
SecondaryAll lanes: 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
Performed under reducing conditions.
Predicted band size: 86 kDa
Observed band size: 80 kDa, 85 kDa, 90 kDa
Western blot - Human DYRK1A knockout HEK-293T cell line (ab267336)
False colour image of Western blot: Anti-DYRK1A antibody staining at 1/500 dilution, 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, the antibody was shown to bind specifically to DYRK1A. A band was observed at 80/95/90 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in DYRK1A knockout cell line ab267336 (knockout cell lysate Human DYRK1A knockout HEK-293T cell lysate ab258402). To generate this image, wild-type and DYRK1A 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
All lanes: Anti-DYRK1A antibody at 1/500 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human DYRK1A knockout HEK-293T cell line (ab267336)
Lane 3: HT1080 cell lysate at 20 µg
Performed under reducing conditions.
Sanger Sequencing - Human DYRK1A knockout HEK-293T cell line (ab267336)
Homozygous: 1 bp insertion in exon12
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