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EIF4EBP1 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: Insertion of the selection cassette in exon 1.

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Images

Western blot - Human EIF4EBP1 knockout HeLa cell line (AB264784), expandable thumbnail
  • Western blot - Human EIF4EBP1 knockout HeLa cell line (AB264784), expandable thumbnail
  • Cell Culture - Human EIF4EBP1 knockout HeLa cell line (AB264784), expandable thumbnail
  • Sanger Sequencing - Human EIF4EBP1 knockout HeLa cell line (AB264784), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1

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

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EIF4EBP1 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: Insertion of the selection cassette in exon 1.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
EIF4EBP1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female

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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HeLa cell line (ab255448). 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, 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

EIF4EBP1 also known as 4EBP1 or p4EBP1 is an important protein that acts as a translational repressor by binding to eIF4E therefore inhibiting cap-dependent translation. The molecular weight of 4EBP1 is approximately 12 kDa. This protein is ubiquitously expressed in various tissues indicating its widespread involvement in cellular functions. eIF4EBP1 can be detected and quantified using techniques like the eIF4EBP1 ELISA making it a common target in research for its role in translation regulation.

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.

Associated diseases and disorders

Deregulation of eIF4EBP1 has been implicated in cancer and neurological disorders. In cancer hyperactive mTOR signaling can lead to excessive phosphorylation of 4EBP1 decreasing its ability to inhibit eIF4E and enhancing translation of oncogenic proteins. In neurological disorders imbalanced eIF4EBP1 activity can disrupt synaptic plasticity and memory formation. Proteins such as p70S6K which are also part of the mTOR pathway share connections with eIF4EBP1 in these pathological conditions.

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