IL13RA2 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.
Images
Key facts
- Cell type
- A-375
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3
Alternative names
CD213a2, Cancer/testis antigen 19, I13R2_HUMAN, IL-13 receptor subunit alpha-2, IL-13R subunit alpha-2, IL-13R-alpha-2, IL-13RA2, IL13 receptor, IL13BP, IL13R, Interleukin 13 receptor alpha 2 chain, Interleukin-13 receptor subunit alpha-2, Interleukin-13-binding protein
Recommended products
IL13RA2 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.
Key facts
- Cell type
- A-375
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3
- Disease
- Melanoma
- Concentration
Properties
- Gene name
- IL13RA2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Immunocytochemistry, Next Generation Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Viability
- ~ 80%
- 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
Recommended control: Human wild-type A375 cell line (ab275461). 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: DMEM (High Glucose) + 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.
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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 IL13RA2 knockout A375 cell line (ab273381)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] ab260044 observed at 49 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] ab260044 was shown to react with IL-13 receptor alpha 2 in wild-type A375 cells in western blot with loss of signal observed in IL13RA2 knockout cell line ab273371 (knockout cell lysate Human IL13RA2 knockout A375 cell lysate ab275532). Wild-type and IL13RA2 knockout A375 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] ab260044 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with 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) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] (Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] ab260044) at 1/1000 dilution
Lane 1: Wild-type A375 cell lysate at 30 µg
Lane 2: IL13RA2 knockout A375 cell lysate at 30 µg
Lane 2: Western blot - Human IL13RA2 knockout A375 cell line (ab273381)
Lane 3: U-87-MG cell lysate at 30 µg
Lane 4: Daudi cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 49 kDa
Immunocytochemistry/ Immunofluorescence - Human IL13RA2 knockout A375 cell line (ab273381)
Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] ab260044 staining IL-13 receptor alpha 2 in wild-type A375 cells (top panel) and IL13RA2 knockout A375 cells (bottom panel) (ab273381). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-IL-13 receptor alpha 2 antibody [EPR22978-163] ab260044 at 1/500 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Next Generation Sequencing - Human IL13RA2 knockout A375 cell line (ab273381)
Homozygous: 1 bp deletion in exon 3
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