HK1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 10.
Images
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 10
Alternative names
BB404130, Brain form hexokinase, DrHXK1, EC 2.7.1.1, Glycolytic enzyme, HEXOKIN, HK I, HK1, HK1-ta, HK1-tb, HK1-tc, HMSNR, HXK1_HUMAN, Hexokinase PI, Hexokinase type I, Hexokinase, tumor isozyme, Hexokinase-1, Hexokinase-A, Hk1-s, dea, hexokinase I, im:7148527, mHk1-s, wu:fc09d08, wu:fc16e02, wu:fc21e02, wu:fq14b11, zgc:55790, zgc:77618
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HK1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 10.
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 10
- Antibiotic resistance
G418 (Geneticin) 1µg/mL
- Concentration
Properties
- Gene name
HK1
- 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 HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
4 product images
Western blot - Human HK1 (Hexokinase 1) knockout HEK-293T cell line (ab267279)
Lanes 1-4: Merged signal (red and green). Green - Anti-Hexokinase 1 antibody [EPR10135(B)] ab154839 observed at 102 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-Hexokinase 1 antibody [EPR10135(B)] ab154839 Anti-Hexokinase 1 antibody [EPR10135(B)] was shown to specifically react with Hexokinase 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267279 (knockout cell lysate Human HK1 (Hexokinase 1) knockout HEK-293T cell lysate ab257161) was used. Wild-type and Hexokinase 1 knockout samples were subjected to SDS-PAGE. Anti-Hexokinase 1 antibody [EPR10135(B)] ab154839 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Hexokinase 1 antibody [EPR10135(B)] (Anti-Hexokinase 1 antibody [EPR10135(B)] ab154839) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HK1 knockout HeLa cell lysate at 20 µg
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 102 kDa
Observed band size: 102 kDa
Western blot - Human HK1 (Hexokinase 1) knockout HEK-293T cell line (ab267279)
Lanes 1-4: Merged signal (red and green). Green - Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker ab150423 observed at 102 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker ab150423 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial was shown to specifically react with Hexokinase 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267279 (knockout cell lysate Human HK1 (Hexokinase 1) knockout HEK-293T cell lysate ab257161) was used. Wild-type and Hexokinase 1 knockout samples were subjected to SDS-PAGE. Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker ab150423 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker ab150423) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HK1 knockout HeLa cell lysate at 20 µg
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: HEPG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 102 kDa
Observed band size: 102 kDa
Cell Culture - Human HK1 (Hexokinase 1) knockout HEK-293T cell line (ab267279)
Representative images of HK1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human HK1 (Hexokinase 1) knockout HEK-293T cell line (ab267279)
Homozygous: 1 bp insertion in exon10
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