Human KRAS knockout HCT116 cell line
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KRAS KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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
CFC2, Cellular c Ki ras2 proto oncogene, Cellular transforming proto oncogene, GTPase KRas, K RAS p21 protein, K RAS2A, K RAS2B, K RAS4A, K RAS4B, K ras, K-Ras 2, KIRSTEN MURINE SARCOMA VIRUS 2, KRAS proto oncogene, GTPase, KRAS1, Ki-Ras, Kirsten rat sarcoma 2 viral (v Ki ras2) oncogene homolog, Kirsten rat sarcoma viral oncogene homolog, N-terminally processed, NS, NS3, Oncogene KRAS2, PR310 c K ras oncogene, RALD, RASK2, RASK_HUMAN, Transforming protein p21, c Ki ras2, c Kirsten ras protein, c-K-ras, c-Ki-ras, p21ras, v Ki ras2 Kirsten rat sarcoma 2 viral oncogene homolog, v Ki ras2 Kirsten rat sarcoma viral oncogene homolog
- WB
Lab
Western blot - Human KRAS knockout HCT116 cell line (AB276083)
Anti-KRAS antibody [EPR23474-76] (ab275876) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab275876 was shown to bind specifically to KRAS. A band was observed at 21 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in KRAS knockout cell line ab276083 (knockout cell lysate ab284205). To generate this image, wild-type and KRAS knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% BSA 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 VeriBlot for IP Detection Reagent (HRP) (ab131366) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Lanes 1 - 2:
Western blot - Anti-KRAS antibody [EPR23474-76] (<a href='/en-us/products/primary-antibodies/kras-antibody-epr23474-76-ab275876'>ab275876</a>) at 1/1000 dilution
Lanes 1 - 2:
Western blot - Anti-KRAS antibody [EPR23474-76] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/kras-antibody-epr23474-76-bsa-and-azide-free-ab275885'>ab275885</a>)
Lane 1:
Wild-type HCT 116 cell lysate at 10 µg
Lane 2:
KRAS knockout HCT 116 cell lysate at 10 µg
Observed band size: 21 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human KRAS knockout HCT116 cell line (AB276083)
Clone A1 sequence : 40 bp deletion
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
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
McCoY5a + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
KRAS influences many cellular processes. It is not part of a complex but acts alone within its pathways. It modulates signal transduction processes which are significant for the control of proliferation apoptosis and cell cycle progress. Mutations within this protein such as KRAS G12V and KRAS G12D lead to continuous activation and can cause uncontrolled cell growth which links them to various cancers. KRAS commonly interacts with proteins like RAF kinase and PI3K signaling components to execute its functions.
Pathways
KRAS is an important component in at least two important signaling pathways: the MAPK/ERK pathway and the PI3K/AKT pathway. These pathways play major roles in regulating gene expression that drives cell growth and division. In the MAPK/ERK pathway KRAS activates RAF which subsequently initiates a phosphorylation cascade that stimulates ERK. This protein also interacts with elements of the PI3K/AKT pathway influencing cellular survival and metabolism. These interactions position KRAS as a vital oncogene in cancer biology.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB108602
BOND RX™ Validated|RabMAb|Recombinant|20ul selling size
![Flow Cytometry (Intracellular) - Anti-Ras antibody [EPR3255] (AB108602)](https://content.abcam.com/products/images/ras-antibody-epr3255-ab108602--flow-cytometry-intracellular-img57702.jpg)
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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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