Human SUZ12 knockout HeLa cell line
- Advanced Validation
- What is this?
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SUZ12 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp deletion in exon 1.
View Alternative Names
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
- WB
Lab
Western blot - Human SUZ12 knockout HeLa cell line (AB264983)
Lanes 1 - 4 : Merged signal (red and green). Green - ab12073 observed at 95 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab12073 was shown to react with SUZ12 in western blot. The band observed in the SUZ12 knockout cell line ab264983 (SUZ12 knockout 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 ab12073 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-SUZ12 antibody (<a href='/en-us/products/primary-antibodies/suz12-antibody-ab12073'>ab12073</a>) 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 2:
Western blot - Human SUZ12 knockout HeLa cell line (ab264983)
Lane 3:
Wild-type HAP1 cell lysate at 20 µg
Lane 4:
SUZ12 knockout HAP1 cell lysate at 20 µg
Predicted band size: 83 kDa
Observed band size: 95 kDa
false
- WB
Lab
Western blot - Human SUZ12 knockout HeLa cell line (AB264983)
Lanes 1- 2 : Merged signal (red and green). Green - ab175187 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
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 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. ab175187 and 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 (ab216773) and 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 (<a href='/en-us/products/primary-antibodies/suz12-antibody-epr5234n-chip-grade-ab175187'>ab175187</a>) 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
Lane 2:
Western blot - Human SUZ12 knockout HeLa cell line (ab264983)
Predicted band size: 83 kDa
Observed band size: 100 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SUZ12 knockout HeLa cell line (AB264983)
Homozygous : 23 bp deletion in exon 1.
Reactivity data
Product details
Recommended control: 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.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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