ARG1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
Hep G2
- Species or organism
Human
- Tissue
Liver
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Alternative names
A I, ARG 1, ARGI1_HUMAN, Al, Arginase liver, Arginase type I, Arginase-1, Liver-type arginase, Type I arginase
Recommended products
ARG1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
Hep G2
- Species or organism
Human
- Tissue
Liver
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Disease
Hepatocellular Carcinoma
- Concentration
Properties
- Gene name
ARG1
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
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
MEM + 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 HepG2 cell line (Human wild-type Hep G2 cell line ab257304). 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:
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3 product images
Western blot - Human ARG1 knockout Hep G2 cell line (ab281603)
False colour image of Western blot: Anti-Liver Arginase antibody [EPR6671(B)] staining at 1/1000 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-Liver Arginase antibody [EPR6671(B)] ab124917 was shown to bind specifically to Liver Arginase. A band was observed at 36 kDa in wild-type HepG2 cell lysates with no signal observed at this size in arg1 knockout cell line ab281603 (knockout cell lysate ab282955). Faint band remaining in KO sample at same molecular weight is likely to be an isoform of arg1. To generate this image wild-type and arg1 knockout HepG2 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-Liver Arginase antibody [EPR6671(B)] (Anti-Liver Arginase antibody [EPR6671(B)] ab124917) at 1/1000 dilution
Lane 1: Wild-type HepG2 cell lysate at 20 µg
Lane 2: arg1 knockout HepG2 cell lysate at 20 µg
Lanes 3 and 5: Empty
Lane 4: Human Liver cell lysate at 5 µg
Lane 6: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 36 kDa
Western blot - Human ARG1 knockout Hep G2 cell line (ab281603)
False colour image of Western blot: Anti-Liver Arginase antibody [EPR6672(B)] staining at 1/1000 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-Liver Arginase antibody [EPR6672(B)] ab133543 was shown to bind specifically to Liver Arginase. A band was observed at 36 kDa in wild-type HepG2 cell lysates with no signal observed at this size in arg1 knockout cell line ab281603 (knockout cell lysate ab282955). To generate this image wild-type and arg1 knockout HepG2 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-Liver Arginase antibody [EPR6672(B)] (Anti-Liver Arginase antibody [EPR6672(B)] ab133543) at 1/1000 dilution
Lane 1: Wild-type HepG2 cell lysate at 20 µg
Lane 2: arg1 knockout HepG2 cell lysate at 20 µg
Lanes 3 and 5: Empty
Lane 4: Human Liver cell lysate at 5 µg
Lane 6: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 35 kDa
Observed band size: 36 kDa
Sanger Sequencing - Human ARG1 knockout Hep G2 cell line (ab281603)
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