Human EIF3K knockout HEK-293T cell line
- Advanced Validation
- What is this?
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EIF3K KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 64 bp deletion in exon 1.
View Alternative Names
ARG134, EIF3K_HUMAN, EIF3S12, Eukaryotic translation initiation factor 3 subunit 12, Eukaryotic translation initiation factor 3 subunit K, HSPC029, M9, MSTP001, Muscle-specific gene M9 protein, PLAC-24, PRO1474, PTD001, eIF-3 p25, eIF-3 p28
- WB
Lab
Western blot - Human EIF3K knockout HEK-293T cell line (AB266841)
Lanes 1-3 : Merged signal (red and green). Green - ab50736 observed at 25 kDa. Red - loading control ab181602 observed at 36 kDa.
ab50736 Anti-eIF3K antibody [2313C2a] was shown to specifically react with eIF3K in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266841 (knockout cell lysate ab258857) was used. Wild-type and eIF3K knockout samples were subjected to SDS-PAGE. ab50736 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-eIF3K antibody [2313C2a] (<a href='/en-us/products/primary-antibodies/eif3k-antibody-2313c2a-ab50736'>ab50736</a>) at 1/500 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
EIF3K knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human EIF3K knockout HEK-293T cell line (ab266841)
Lane 3:
THP1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human EIF3K knockout HEK-293T cell line (AB266841)
Homozygous : 64 bp deletion in exon 1
- Cell Culture
Lab
Cell Culture - Human EIF3K knockout HEK-293T cell line (AB266841)
Representative images EIF3K knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T 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 M5000 microscope.
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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.
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
EIF3K is involved in the regulation of the eukaryotic translation initiation factor 3 complex which is essential for initiating translation by serving as a scaffold for other eIF3 subunits. This complex stabilizes the formation of the pre-initiation complex necessary for ribosome binding to mRNA. Through its involvement in protein synthesis eIF3K influences cell growth proliferation and response to various cellular signals. It is involved in processes such as cell cycle control and apoptotic regulation as it aids the translation of specific mRNAs coding for key regulatory proteins.
Pathways
EIF3K participates in mTOR and Wnt signaling pathways. These pathways are critical in controlling cell growth proliferation and differentiation. Through mTOR signaling eIF3K interacts with proteins like ribosomal protein S6 kinase beta-1 (S6K1) influencing cell metabolism and growth. In the Wnt pathway eIF3K contributes to the translation of proteins that regulate cell fate and stem cell renewal. Interactions with these pathways illustrate the importance of eIF3K in maintaining cellular homeostasis.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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