Human NRAS knockout HEK-293T cell line
- Advanced Validation
- What is this?
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NRAS KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon 2 and 8 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
ALPS4, AV095280, GTPase NRas, H-Ras-1, N ras, N ras protein part 4, NRAS1, NS6, Neuroblastoma RAS viral (v ras) oncogene homolog, OTTHUMP00000013879, OTTMUSP00000023521, RASN_HUMAN, Transforming protein N-Ras, v ras neuroblastoma RAS viral oncogene homolog
- WB
Lab
Western blot - Human NRAS knockout HEK-293T cell line (AB266684)
Lanes 1 - 4 : Merged signal (red and green). Green - ab198820 observed at 22 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab198820 was shown to react with NRAS in wild-type HEK-293T cells in Western blot with loss of signal observed in NRAS knockout cell line ab266684 (NRAS knockout cell lysate ab258542). Wild-type HEK-293T and NRAS knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab198820 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 200 dilution 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-NRAS antibody - C-terminal (<a href='/en-us/products/primary-antibodies/nras-antibody-c-terminal-ab198820'>ab198820</a>) at 1/200 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
NRAS knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human NRAS knockout HEK-293T cell line (ab266684)
Lane 3:
MCF7 cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Predicted band size: 21 kDa
Observed band size: 22 kDa
false
- WB
Lab
Western blot - Human NRAS knockout HEK-293T cell line (AB266684)
Lanes 1 - 4 : Merged signal (red and green). Green - ab167136 observed at 22 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab167136 was shown to react with NRAS in wild-type HEK-293T cells in Western blot with loss of signal observed in NRAS knockout cell line ab266684 (NRAS knockout cell lysate ab258542). Wild-type HEK-293T and NRAS knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab167136 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 0.5 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-NRAS antibody (<a href='/en-us/products/primary-antibodies/nras-antibody-ab167136'>ab167136</a>) at 0.5 µg/mL
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
NRAS knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human NRAS knockout HEK-293T cell line (ab266684)
Lane 3:
MCF7 cell lysate at 20 µg
Lane 4:
A549 cell lysate at 20 µg
Predicted band size: 21 kDa
Observed band size: 22 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human NRAS knockout HEK-293T cell line (AB266684)
Allele-2 : 5 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human NRAS knockout HEK-293T cell line (AB266684)
Allele-1 : 8 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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
NRAS plays an important role in cell proliferation differentiation and survival. It does not function alone but is part of the larger RAS protein family including HRAS and KRAS with whom it shares similar roles and sequence homology. NRAS interacts with various growth factor receptors and mediates signals to downstream effectors that influence cellular processes. Mutations in the NRAS gene can affect these biological processes leading to altered cell behavior.
Pathways
NRAS has significant roles in the MAPK/ERK pathway and the PI3K/AKT pathway. These pathways regulate fundamental cellular functions including growth and survival. NRAS interacts closely with other proteins such as RAF kinases and PI3K to propagate signals from activated receptors at the cell membrane to the nucleus. Altered NRAS activity can impact these pathways affecting cellular responses to external stimuli.
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
Complementary Products
Primary Antibodies
AB227658
Anti-Ras (mutated Q61R) antibody [SP174]
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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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