SUZ12 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp deletion in exon 1.
HeLa
Human
Cervix
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp deletion in exon 1
CHET9, ChET 9 protein, Chromatin precipitated E2F target 9 protein, JJAZ1, Joined to JAZF1 protein, KIAA0160, Polycomb protein SUZ12, SUZ12 polycomb repressive complex 2 subunit, SUZ12_HUMAN, Suppressor of zeste 12 homolog, Suppressor of zeste 12 protein homolog
SUZ12 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp deletion in exon 1.
HeLa
Human
Cervix
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp deletion in exon 1
Puromycin 1µg/mL
Adenocarcinoma
SUZ12
Knockout
CRISPR technology
Sanger Sequencing
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). 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.
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.
Lanes 1 - 4: Merged signal (red and green). Green - Anti-SUZ12 antibody ab12073 observed at 95 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
Anti-SUZ12 antibody ab12073 was shown to react with SUZ12 in western blot. The band observed in the SUZ12 knockout cell line ab264983 (SUZ12 knockout cell lysate Human SUZ12 knockout HeLa cell lysate ab257721) lane below 95 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-SUZ12 antibody ab12073 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at 1 μg/ml 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 h at room temperature before imaging.
All lanes: Western blot - Anti-SUZ12 antibody (Anti-SUZ12 antibody ab12073) at 1 µg/mL
Lane 1: Wild-type Hela cell lysate at 20 µg
Lane 2: SUZ12 knockout HeLa cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: SUZ12 knockout HAP1 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 95 kDa
Lanes 1- 2: Merged signal (red and green). Green - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 was shown to react with SUZ12 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264983 (CRISPR/Cas9 edited cell lysate Human SUZ12 knockout HeLa cell lysate ab257721) lane below 100kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and SUZ12 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade ab175187) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: SUZ12 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 83 kDa
Observed band size: 100 kDa
Homozygous: 23 bp deletion in exon 1.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com