Human EIF4EBP1 knockout HeLa cell line
- Advanced Validation
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EIF4EBP1 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: Insertion of the selection cassette in exon 1. 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
4EBP1_HUMAN, BP 1, Eif4ebp1, Eukaryotic translation initiation factor 4E-binding protein 1, PHAS-I, Phosphorylated heat- and acid-stable protein regulated by insulin 1, eIF4E-binding protein 1
- WB
Lab
Western blot - Human EIF4EBP1 knockout HeLa cell line (AB264784)
Lane 1 : Wild-type HeLa cell lysate (20 µg)
Lane 2 : EIF4EBP1 knockout HeLa cell lysate (20 µg)
Lanes 1-2 : Merged signal (red and green). Green - ab32024 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32024 Anti-eIF4EBP1 antibody [Y329] was shown to specifically react with eIF4EBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32024 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-eIF4EBP1 antibody [Y329] (<a href='/en-us/products/primary-antibodies/eif4ebp1-antibody-y329-ab32024'>ab32024</a>) at 1/5000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EIF4EBP1 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-eif4ebp1-knockout-hela-cell-lysate-ab257146'>ab257146</a>) at 20 µg
Secondary
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Lanes 1 - 3:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 13 kDa
Observed band size: 13 kDa,37 kDa
false
- WB
Lab
Western blot - Human EIF4EBP1 knockout HeLa cell line (AB264784)
Lanes 1-2 : Merged signal (red and green). Green - ab32130 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32130 Anti-eIF4EBP1 antibody [Y330] was shown to specifically react with eIF4EBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-eIF4EBP1 antibody [Y330] (<a href='/en-us/products/primary-antibodies/eif4ebp1-antibody-y330-ab32130'>ab32130</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
EIF4EBP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EIF4EBP1 knockout HeLa cell line (ab264784)
Predicted band size: 13 kDa,36 kDa
Observed band size: 13 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human EIF4EBP1 knockout HeLa cell line (AB264784)
Homozygous : Insertion of the selection cassette in exon 1.
- Cell Culture
Unknown
Cell Culture - Human EIF4EBP1 knockout HeLa cell line (AB264784)
Representative images of EIF4EBP1 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.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
EIF4EBP1 plays an important role in regulating cell growth and proliferation by modulating protein synthesis. It is a part of the eIF4F complex which is responsible for the initiation of mRNA translation. When hypophosphorylated eIF4EBP1 binds tightly to eIF4E and prevents the assembly of the active eIF4F complex leading to reduced translation initiation of mRNAs involved in growth and survival. This regulation is important in conditions where cells need to adapt to metabolic stress or external signals.
Pathways
The mammalian target of rapamycin (mTOR) pathway regulates eIF4EBP1 through phosphorylation. In response to growth signals mTOR phosphorylates eIF4EBP1 causing the release of eIF4E and allowing mRNA translation to proceed. This interaction links eIF4EBP1 to the PI3K/AKT/mTOR signaling pathway which influences cell cycle progression and survival. Related proteins in this pathway include ribosomal protein S6 kinase (S6K1) which is also phosphorylated by mTOR to promote protein synthesis.
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
AB32024
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated
![Flow Cytometry (Intracellular) - Anti-eIF4EBP1 antibody [Y329] (AB32024)](https://content.abcam.com/products/images/eif4ebp1-antibody-y329-ab32024--flow-cytometry-intracellular-img68578.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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