CASP9 KO cell line available to order. Free of charge wild type control provided.
Images
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
Alternative names
APAF-3, Apoptosis related cysteine peptidase, Apoptotic protease Mch-6, Apoptotic protease-activating factor 3, CASP9_HUMAN, Caspase 9 Dominant Negative, Caspase 9 apoptosis related cysteine peptidase, Caspase 9c, Caspase-9, Caspase-9 subunit p10, ICE-LAP6, ICE-like apoptotic protease 6, MCH6, PPP1R56, RNCASP9, protein phosphatase 1, regulatory subunit 56
Recommended products
CASP9 KO cell line available to order. Free of charge wild type control provided.
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Disease
- Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
- CASP9
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Suspension
- 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. 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.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
- Culture medium
- RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
- 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 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:
Terms & Conditions.
3 product images
Western blot - Human CASP9 knockout THP-1 cell line (ab276122)
False colour image of Western blot: Anti-Caspase-9 antibody [E23] staining at 1/1000 dilution shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-Caspase-9 antibody [E23] ab32539 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image wild-type and CASP9 knockout THP-1 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-Caspase-9 antibody [E23] (Anti-Caspase-9 antibody [E23] ab32539) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: CASP9 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human CASP9 knockout THP-1 cell line (ab276122)
Lane 3: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 45 kDa
Western blot - Human CASP9 knockout THP-1 cell line (ab276122)
False colour image of Western blot: Anti-Caspase-9 antibody [EPR18107] staining at 1/2000 dilution shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-Caspase-9 antibody [EPR18107] ab202068 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image wild-type and CASP9 knockout THP-1 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-Caspase-9 antibody [EPR18107] (Anti-Caspase-9 antibody [EPR18107] ab202068) at 1/2000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: CASP9 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human CASP9 knockout THP-1 cell line (ab276122)
Lane 3: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 45 kDa
Western blot - Human CASP9 knockout THP-1 cell line (ab276122)
False colour image of Western blot: Anti-Caspase-9 antibody [EPR18108] staining at 1/1000 dilution shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-Caspase-9 antibody [EPR18108] ab185719 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line ab276122 (knockout cell lysate ab284219). To generate this image wild-type and CASP9 knockout THP-1 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-Caspase-9 antibody [EPR18108] (Anti-Caspase-9 antibody [EPR18108] ab185719) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: CASP9 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human CASP9 knockout THP-1 cell line (ab276122)
Lane 3: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 45 kDa
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