EIF4EBP1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
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
EIF4EBP1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
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.
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.
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.
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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Lane 2: EIF4EBP1 knockout HeLa cell lysate (20 μg)
Lanes 1-2: Merged signal (red and green). Green - Anti-eIF4EBP1 antibody [Y330] ab32130 observed at 13 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-eIF4EBP1 antibody [Y330] 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 Human EIF4EBP1 knockout HeLa cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. Anti-eIF4EBP1 antibody [Y330] ab32130 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-eIF4EBP1 antibody [Y330] ab32130) 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 (Human EIF4EBP1 knockout HeLa cell line ab264784)
Performed under reducing conditions.
Predicted band size: 13 kDa, 36 kDa
Observed band size: 13 kDa
Lane 2: EIF4EBP1 knockout HeLa cell lysate (20 μg)
Lanes 1-2: Merged signal (red and green). Green - Anti-eIF4EBP1 antibody [Y329] ab32024 observed at 13 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-eIF4EBP1 antibody [Y329] 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 Human EIF4EBP1 knockout HeLa cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. Anti-eIF4EBP1 antibody [Y329] ab32024 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-eIF4EBP1 antibody [Y329] ab32024) at 1/5000 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 (Human EIF4EBP1 knockout HeLa cell line ab264784)
Performed under reducing conditions.
Predicted band size: 13 kDa
Observed band size: 13 kDa
Homozygous: Insertion of the selection cassette in exon 1
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