KRAS KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
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
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KRAS KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- KRAS
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The KRAS protein also known as Kirsten rat sarcoma viral oncogene homolog is a small GTPase weighing around 21 kDa. It plays an important role in cell signaling pathways and is widely expressed in various tissues. As a molecular switch KRAS cycles between an active GTP-bound state and an inactive GDP-bound state regulating cell proliferation differentiation and survival. KRAS mutations are often studied using various cell lines including the HCT116 line which is a human colorectal carcinoma cell line with a KRAS mutant background. Western blot analysis commonly helps in investigating KRAS protein expression and activity levels.
- 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.
- Associated diseases and disorders
KRAS mutations are important in the development and progression of colorectal and pancreatic cancers. The persistence of active KRAS leads to continuous signaling that drives tumor growth and resistance to apoptosis. KRAS-associated cancers often show poor prognosis partly due to limited therapeutic options. Its interaction with proteins like EGFR (Epidermal Growth Factor Receptor) in various cancers highlights complex treatment challenges as these proteins can provide alternative signaling routes further complicating disease management.
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2 product images
Western blot - Human KRAS knockout HCT116 cell line (ab276083)
Anti-KRAS antibody [EPR23474-76] (Anti-KRAS antibody [EPR23474-76] ab275876) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-KRAS antibody [EPR23474-76] 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) (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] (Anti-KRAS antibody [EPR23474-76] ab275876) at 1/1000 dilution
Lanes 1 - 2: Western blot - Anti-KRAS antibody [EPR23474-76] - BSA and Azide free (Anti-KRAS antibody [EPR23474-76] - BSA and Azide free ab275885)
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
Sanger Sequencing - Human KRAS knockout HCT116 cell line (ab276083)
Clone A1 sequence: 40 bp deletion
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