Human MET (Met (c-Met)) knockout HeLa cell line
- Advanced Validation
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(1 Publication)
MET KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
AUTS9, D249, HGF receptor, HGF/SF receptor, HGFR, Hepatocyte growth factor receptor, MET proto oncogene, receptor tyrosine kinase, MET_HUMAN, Met proto oncogene tyrosine kinase, Met proto-oncogene, Met proto-oncogene (hepatocyte growth factor receptor), Oncogene MET, Proto-oncogene c-Met, RCCP2, SF receptor, Scatter factor receptor, Tyrosine-protein kinase Met, c met
- WB
Lab
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Western blot : Anti-MET antibody [EP1454Y] (ab51067) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab51067 was shown to bind specifically to MET. A band was observed at 40-50 kDa in wild-type A549 cell lysates with no signal observed at this size in MET knockout cell line. To generate this image, wild-type and MET knockout A549 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°C. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Lanes 1 - 6:
Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-ep1454y-n-terminal-ab51067'>ab51067</a>) at 1/1000 dilution
Lanes 1 - 2:
Western blot - Anti-IMPDH2 antibody [EPR8365(B)] (<a href='/en-us/products/primary-antibodies/impdh2-antibody-epr8365b-ab129165'>ab129165</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
MET knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HeLa ab255929 cell lysate at 20 µg
Lane 4:
MET (Met (c-Met)) knockout HeLa <a href='/en-us/products/cell-lysates/human-met-c-met-knockout-hela-cell-lysate-ab256991'>ab256991</a> cell lysate at 20 µg
Lane 5:
HT-29 cell lysate at 20 µg
Lane 6:
T-47D cell lysate at 20 µg
Secondary
All lanes:
NFDM/TBST at 1/20000 dilution
Observed band size: 40-50 kDa
false
- WB
Lab
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Western blot : Anti-MET antibody [EPR19554-110] (ab254252) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab254252 was shown to bind specifically to MET. A band was observed at 80-140 kDa in wild-type A549 cell lysates with no signal observed at this size in MET knockout cell line. To generate this image, wild-type and MET knockout A549 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°C. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. The full length MET protein is glycosylated and has multiple cleavage sites, so cleaved products can be detected between ~40-140 kDa
All lanes:
Western blot - Anti-Met (c-Met) antibody [EPR19554-110] (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-epr19554-110-ab254252'>ab254252</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
MET knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HeLa ab255929 cell lysate at 20 µg
Lane 4:
MET (Met (c-Met)) knockout HeLa <a href='/en-us/products/cell-lysates/human-met-c-met-knockout-hela-cell-lysate-ab256991'>ab256991</a> cell lysate at 20 µg
Lane 5:
HT-29 cell lysate at 20 µg
Lane 6:
T-47D cell lysate at 20 µg
Observed band size: 80-140 kDa
true
- Cell Culture
Unknown
Cell Culture - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Representative images of MET knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Homozygous : 5 bp deletion in exon 2.
Reactivity data
Product details
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.
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
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
eLife 12: PubMed39268701
2024
Applications
Unspecified application
Species
Unspecified reactive species
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Product promise
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