MKI67 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 7 bp deletion in exon 7.
Images
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 7 bp deletion in exon 7
Alternative names
Antigen KI-67, Antigen identified by monoclonal antibody Ki-67, KI67_HUMAN, KIA, MKI67, Marker of proliferation Ki-67, PPP1R105, Proliferation marker protein Ki-67, Proliferation related Ki 67 antigen, Protein phosphatase 1 regulatory subunit 105, RP11-380J17.2
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MKI67 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 7 bp deletion in exon 7.
Alternative names
Antigen KI-67, Antigen identified by monoclonal antibody Ki-67, KI67_HUMAN, KIA, MKI67, Marker of proliferation Ki-67, PPP1R105, Proliferation marker protein Ki-67, Proliferation related Ki 67 antigen, Protein phosphatase 1 regulatory subunit 105, RP11-380J17.2
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 7 bp deletion in exon 7
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
MKI67
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
- Zygosity
Homozygous
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- 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 long-term storage conditions
-196°C
Notes
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.
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.
2 product images
Western blot - Human MKI67 (Ki67) knockout HeLa cell line (ab255407)
Lanes 1 - 3: Merged signal (red and green). Green - Anti-Ki67 antibody [SP6] ab16667 observed at 359 kDa. Red - loading control Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.
Anti-Ki67 antibody [SP6] ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate Human MKI67 (Ki67) knockout HeLa cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. Anti-Ki67 antibody [SP6] ab16667 and Anti-Vinculin antibody [VIN-54] (Anti-Vinculin antibody [VIN-54] ab130007) were incubated overnight at 4° at 1 in 100 dilution and 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.
Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) at 1/100 dilution
Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6] - BSA free (Anti-Ki67 antibody [SP6] - BSA free ab231172) at 1/100 dilution
Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6], prediluted (Anti-Ki67 antibody [SP6], prediluted ab21700) at 1/100 dilution
Lane 1: Ramos cell lysate at 20 µg
Lane 2: Wild-type HeLa cell lysate at 20 µg
Lane 3: MKI67 knockout HeLa cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 358 kDa
Observed band size: 124 kDa, 359 kDa
Lanes 1 - 3: Merged signal (red and green). Green - Anti-Ki67 antibody [SP6] ab16667 observed at 359 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.
Anti-Ki67 antibody [SP6] ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate Human MKI67 (Ki67) knockout HeLa cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. Anti-Ki67 antibody [SP6] ab16667 and Anti-Vinculin antibody [VIN-54] (Anti-Vinculin antibody [VIN-54] ab130007) were incubated overnight at 4°C at 1 in 100 dilution and 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.
Sanger Sequencing - Human MKI67 (Ki67) knockout HeLa cell line (ab255407)
Homozygous: 7 bp deletion in exon 7.
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